{"id":54427,"date":"2023-12-31T10:04:29","date_gmt":"2023-12-31T10:04:29","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=54427"},"modified":"2024-01-05T08:02:28","modified_gmt":"2024-01-05T08:02:28","slug":"antioxidant-activity-and-phytochemical-profile-of-diadema-paucispinum-from-sumenep-madura-indonesia","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol16no4\/antioxidant-activity-and-phytochemical-profile-of-diadema-paucispinum-from-sumenep-madura-indonesia\/","title":{"rendered":"Antioxidant Activity and Phytochemical Profile of Diadema paucispinum from Sumenep-Madura, Indonesia"},"content":{"rendered":"\n

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

Sea urchin, locally known as sea urchin\/tehe-tehe, is a type of Echinodermata phylum, Echinoidea class, with a round shape covered with long moving spines 1<\/sup>. Sea urchins act as bioindicators in controlling vegetation growth in the sea because their leading food is seaweed and microalgae 2<\/sup>. Sea urchins are nocturnal foragers, while during the day, they hide in coral crevices 3<\/sup>. Diadema paucispinum<\/em> is found in the Sumenep-Madura Sea, and is not utilized by fishermen, so that many sea urchin habitats die in the waters around the coast and become marine litter. <\/p>\n\n\n\n

Japan is one of the countries that utilize sea urchin as a potential food due to the high protein content in its gonads. The utilization of the shell has yet to be widely studied and is only thrown away, even though the sea urchin shell contains pigment compounds, including PHNQ, which has the potential as a high antioxidant and potential as a cosmetic ingredient 4<\/sup>. These pigment compounds vary in sea urchins. It depends on the habitat conditions in which they live 5<\/sup>. The darker or more colorful the shell pigments, the more diverse the absorption of chemical compounds and the higher the antioxidant power 6,7<\/sup>, so exploring the metabolite content of sea urchin shells based on their habitat is essential. Using sea urchin shells and gonads will increase their antioxidant activity 7<\/sup>.<\/p>\n\n\n\n

Sea urchins have a hard shell and contain gonads containing essential amino acids, \u00df-carotene, and DHA [8;9]. In the analysis of Diadema Sp<\/em> gonads, 80% polyunsaturated fatty acids (PUFA) were found, such as eicosapentaenoic acid (EPA), and other components like arachidonic acid, carotenoids, and containing antioxidant compounds, such as echinenone, \u00df-carotene and fucoxanthin 10,11<\/sup>. Nutrients in sea urchin gonads include vitamins A, B9, PUFAs, and flavonoids, predicted suppress chronic inflammation 12<\/sup>. Sea urchin shells contain bioactive compounds known as polyhydroxy naphthoquinone (PHNQ) derivatives such as Spinochrome A, B, C, D, E, Echinocrome, and Echinamine A, B, E. PHNQ in the shells is a pigmen specific compound in sea urchin, proves to have pharmacological potential actions as antioxidants and anti-inflammatory agent 13<\/sup>.<\/p>\n\n\n\n

Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS) are the most widely recognized free radicals. ROS mediates intracellular damage to lipids, proteins, carbohydrates, and nucleic acids. ROS are highly reactive because they are unstable (have unpaired electrons). Oxidative stress occurs when an imbalance between oxidant and antioxidant molecules increases ROS production, resulting in tissue damage and inflammation. Antioxidant compounds can slow down\/prevent the oxidation process that produces free radicals and break down chains that can damage cells and tissues, so antioxidants can be used as a therapeutic option for inflammation, which is generally caused by ROS 14<\/sup>.<\/p>\n\n\n\n

Inflammatory processes can arise acutely or chronically, cause local or systemic impacts, and cause pathological abnormalities. The primary treatment for inflammation is to relieve pain and stop tissue damage. The drugs given are steroids and non-steroidal anti-inflammatory drugs (NSAIDs), which have serious side effects, such as an increased risk of gastric ulcers and upper gastrointestinal bleeding 15<\/sup>.<\/p>\n\n\n\n

Several studies derived from marine biota on anti-inflammatory agents also have strong antioxidant potential. Sea urchin is a marine biota with anti-inflammatory, antioxidant, and antidiabetic activities 3<\/sup>. The anti-inflammatory activity of methanol extract of the sea urchin Echinometra mathaei<\/em> showed high anti-inflammatory activity 16<\/sup>. In the 70% ethanol extract of Echinometra mathaei<\/em>, one type of sea urchin from Weh Island, Sabang, proved to be a more potent anti-inflammatory than diclofenac sodium at a dose of 100 mg\/KgBB 17<\/sup>.<\/p>\n\n\n\n

Research on Diadema fauscispinum<\/em> itself is mostly just its morphological, anatomical, and phylogenetic properties. Activity testing and active metabolite content of this species have not been found, primarily based on the closeness of its genus, Diadema setosum<\/em>. Based on the closeness of the genus, Diadema sp.<\/em> known has PHNQ content in both the shell and gonads 18<\/sup>. The more researched type of long-spine black sea urchin is Diadema setosum<\/em>, which has been studied for its gonad extract as a burn agent in rabbits and has the potential for 100% burn healing in 1 to 2-degree burns 19<\/sup>. The potential of Diadema sp<\/em> as a burn medicine is related to its anti-inflammatory properties. Diadema sp<\/em> research in Indonesia states that this sea urchin is proven to have antibacterial activity 20,21<\/sup>, reduce IFN, increase IL-10 22<\/sup>, have antioxidant activity 23,24<\/sup>, have flavonoids, steroids, saponins 24<\/sup>, and do not cause toxicity in the BSLT test 25<\/sup>. Diadema research in Egypt and Vietnam found phenolic content, PHNQ, antioxidant activity, and antibacterial properties 4,26:27<\/sup>. In addition, Diadema sp<\/em> research shows low toxicity26,28<\/sup>. Diadema fauscispinum <\/em>was choosen in this study because Diadema<\/em> species is the most found species in Indonesian coral seas. It will be utilized as much as possible for health and beauty. The antioxidant potential of sea urchin is excellent and has potential for development in various treatments; besides, its antioxidant power is related to its potential as an essential ingredient for cosmetic preparations.<\/p>\n\n\n\n

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

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

Preparation of research test materials\nincludes drying shells (simplicia) and extraction with 96% ethanol. The ethanol\nextract of Diadema paucispinum<\/em> was prepared by maceration using 96%\nethanol; the filtrate was collected and evaporated with a rotary vacuum\nevaporator to obtain a thick extract. The percentage yield of the extract was\ncalculated against the weight of the initial simplicia used. The extract’s\nphytochemical screening and FTIR analysis identified the secondary metabolite\nprofile.<\/p>\n\n\n\n

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

Aquadest, 96% ethanol, FeCl3, Mg powder, HClconct,\nDragendroff reagent, Mayer reagent, acetic acid anhydrous, H2<\/sub>SO4<\/sub>,\nand Libermann-Burchard \nreagent.<\/p>\n\n\n\n

Phytochemical Screening<\/strong><\/p>\n\n\n\n

Phytochemical Screening test: 3 g sample extract, reagent, and opinions. The sample extract is then subjected to multiple operations and observed for color changes, precipitation, or other phenomena as directed by the process, the test based on Shaikh and Patil research 29<\/sup>.<\/p>\n\n\n\n

Infrared Spectroscopy Profile\nAnalysis <\/strong><\/p>\n\n\n\n

Infrared spectroscopy profile analysis takes\nsamples of 0.5-1.5 mg of chemicals inserted into the sample holder, then scans\nusing Infrared spectroscopy.<\/p>\n\n\n\n

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

Preparation of DPPH\nreagent.<\/strong><\/p>\n\n\n\n

Weigh 4 mg of DPPH dissolved with methanol to\n100 ml (40 ppm).<\/p>\n\n\n\n

Preparation of ABTS reagent<\/strong><\/p>\n\n\n\n

Reagent 1. A 7 mM ABTS solution was made by\nweighing 19.2045 mg of ABTS dissolved in 5 ml of distilled water.<\/p>\n\n\n\n

Reagent 2. Make a 2.45 mM potassium\npersulphate solution by weighing 3.31 mg of potassium persulphate in 5 ml of\ndistilled water. Mix 5 ml of reagent 1 and 5 ml of reagent 2,  and incubate in a dark room for 12-16 hours.\nDissolve the ABTS reagent mixture with distilled water until the absorbance\nvalue is about 0.7 at a wavelength of 734 nm.<\/p>\n\n\n\n

Preparation of\nvitamin C standard<\/strong><\/p>\n\n\n\n

Prepare a concentration series by diluting a\nsolution of 10 ppm, 20 ppm, 30 ppm, and 40 ppm. Measurement with a\nspectrophotometer is done by mixing each sample concentration series as much as\n0.1 ml plus 0.9 ml ABTS reagent \/ DPPH reagent, incubating for 6 minutes, then\nmeasuring the absorbance at a wavelength of 734 nm.<\/p>\n\n\n\n

Sea urchin (Diadema paucispinum<\/em>) sample preparation<\/strong><\/p>\n\n\n\n

Prepare a master solution with a\nconcentration of 50,000 ppm by weighing 253.5 mg of sea urchin sample dissolved\nwith 96% ethanol to a volume of 5 ml. A concentration series was made by\ndiluting the previous solution into concentrations of 5.070 ppm, 10.140 ppm,\n15.210 ppm, 20.280 ppm, 25.350 ppm, 30.420 ppm, and 40.560 ppm.<\/p>\n\n\n\n

Measurement of\nDPPH\/ABTS Binding Activity<\/strong><\/p>\n\n\n\n

Measurement with a spectrophotometer was\ncarried out by mixing each sample concentration series of 0.2 ml plus 0.8 ml\nDPPH\/ABTS reagent, incubated for 30 minutes, then measuring the absorbance at a\nwavelength of 515 nm for DPPH and 734 nm for ABTS.<\/p>\n\n\n\n

The antioxidant activity is measured based on the formula:<\/p>\n\n\n\n

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

Data analysis of IC50<\/sub> DPPH\/ABTS\ninhibition against extract concentration was calculated based on the regression\nequation of % inhibition.
\n<\/strong><\/p>\n\n\n\n

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

Determination results of sea urchin (Diadema paucispinum<\/em>)<\/strong><\/p>\n\n\n\n

Sea urchins were determined at the Biology\nService Unit, Faculty of Science and Technology, Universitas Airlangga, East\nJava. Determination results showed that the samples used were sea urchins with\nthe species Diadema paucispinum<\/em>. The following is the classification of Diadema\npaucispinum<\/em> sea urchin we found.<\/p>\n\n\n\n

Kingdom       :\nAnimalia<\/em><\/p>\n\n\n\n

Phylum         : Echinodermata<\/em><\/p>\n\n\n\n

Subphylum   : Echinozoa<\/em><\/p>\n\n\n\n

Class              : Echinoidea<\/em><\/p>\n\n\n\n

Subclass       : Euechinoidea<\/em><\/p>\n\n\n\n

Infraclass     : Aulodonta<\/em><\/p>\n\n\n\n

Superorder   : Diadematacea<\/em><\/p>\n\n\n\n

Order            : Diadematoida<\/em><\/p>\n\n\n\n

Family           : Diadematidae<\/em><\/p>\n\n\n\n

Genus           : Diadema<\/em><\/p>\n\n\n\n

Species         : Diadema paucispinum <\/em><\/p>\n\n\n\n

Synonyms     : Centrechinus paucispinus <\/em><\/p>\n\n\n\n

Macroscopic\nTesting Results<\/strong><\/p>\n\n\n\n

In the macroscopic test, observations are made by looking at the physical appearance to determine the characteristics of sea urchins (Diadema paucispinum<\/em>). The results of the macroscopic test determination of sea urchin (Diadema paucispinum<\/em>) are shown in the figure and table below.<\/p>\n\n\n\n\n\n
\"\"<\/td>\nFigure 1: Sea urchin (Diadema paucispinum).<\/em><\/strong>

<\/p>\n

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

Table 1: Macroscopic test results of sea urchin (Diadema paucispinum)<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

Parameters<\/p>\n<\/td>\n

\n

Observation results<\/p>\n<\/td>\n

\n

Reff Standard *<\/p>\n<\/td>\n<\/tr>\n

\n

Shape<\/p>\n

Color<\/p>\n

Size<\/p>\n<\/td>\n

\n

Oval round<\/p>\n

Brownish black<\/p>\n

Width : 7,3 cm<\/p>\n

Length : 7,8 cm<\/p>\n

Height : 4,5 cm<\/p>\n<\/td>\n

\n

–<\/p>\n

–<\/p>\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Requirement (*) = Standard reference not found<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Microscopic\nTesting Results<\/strong><\/p>\n\n\n\n

The following results are microscopic\nobservations of sea urchin (Diadema paucispinum<\/em>) shells simplicial.\nMicroscopic observations in chloralhydrate under 100 times magnification showed\nmany prism-shaped calcium oxalate crystals, needle-shaped calcium oxalate\ncrystals, and more-shaped calcium oxalate crystals. It was also found in oil\ncells and trichomes<\/p>\n\n\n\n

Extraction Results of Sea urchin (<\/strong>Diadema paucispinum<\/strong>)<\/strong><\/p>\n\n\n\n

Sea urchin (Diadema paucispinum) was extracted using maceration\nmethod with 96% ethanol solvent.  The\nextraction results obtained a yield of 4.87% which is the amount of compounds\nthat are attracted during the extraction process using 96% ethanol solvent. The\nresulting yield is relatively smaller at 4.87% of the dry weight of 600g. The\nhigher the percent yield value obtained, the more extracts obtained. <\/p>\n\n\n\n

Phytochemical\nScreening Test Results <\/strong><\/p>\n\n\n\n

The following is a phytochemical\nscreening test of 96% ethanol extract of sea urchin (Diadema paucispinum) shown in the table below.<\/p>\n\n\n\n

Table 2: Phytochemical Screening Results of 96% Ethanol Extract Diadema paucispinum<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

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

\n

Test *)<\/sup><\/strong><\/p>\n<\/td>\n

\n

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

\n

Results <\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Alkaloid<\/p>\n<\/td>\n

\n

Mayer reagens<\/p>\n<\/td>\n

\n

Mayers test<\/p>\n<\/td>\n

\n

Forms a yellowish-white precipitate<\/p>\n<\/td>\n

\n

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

\n

Flavonoid<\/p>\n<\/td>\n

\n

Extract + methanol +<\/p>\n

Mg + HCl conc.<\/p>\n<\/td>\n

\n

Shibata\u2019s test<\/p>\n<\/td>\n

\n

Form dark red colour<\/p>\n

(flavonols)<\/p>\n<\/td>\n

\n

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

\n

Saponin<\/p>\n<\/td>\n

\n

Hot water<\/p>\n<\/td>\n

\n

Foaming test<\/p>\n<\/td>\n

\n

foaming form 1,5 cm height<\/p>\n<\/td>\n

\n

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

\n

Tannins<\/p>\n<\/td>\n

\n

FeCl3<\/sub> 5%<\/p>\n<\/td>\n

\n

Braymer\u2019s test<\/p>\n<\/td>\n

\n

Blue-Greenish ring<\/p>\n<\/td>\n

\n

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

\n

Terpenoid<\/p>\n

 <\/p>\n<\/td>\n

\n

0.5 mL Chloroform + 0.5 mL anhydrous acetic acid + 1-2 mL concentrated sulfuric acid (H2SO4)<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n

\n

A red rose colour : Not formed<\/p>\n<\/td>\n

\n

–<\/p>\n<\/td>\n<\/tr>\n

\n

Steroid<\/p>\n<\/td>\n

\n

Anhydrous acetic acid + concentrated sulphate (H2SO4)<\/p>\n<\/td>\n

\n

Libermann-Burchard test<\/p>\n<\/td>\n

\n

An array of colour change: Not formed<\/p>\n<\/td>\n

\n

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

* literature based on Shaikh and Patil (2020)<\/p>\n\n\n

The table above shows the results of the phytochemical screening test of 96% ethanol extract of sea urchin (Diadema paucispinum).<\/em> Phytochemical screening tests include steroids, tannins, flavonoids, saponins, alkaloids, and terpenoids. The results of the chemical content test showed that 96% ethanol extract of sea urchin (Diadema paucispinum<\/em>) positively contained alkaloid compounds, flavonoids, saponins, and tannins and negatively contained steroid compounds, and terpenoids.<\/p>\n\n\n\n

Results of FTIR Analysis of Sea Urchin (Diadema paucispinum<\/em>)<\/strong><\/p>\n\n\n\n

Samples\nof sea urchin were obtained, and simplicia and extracts with FTIR were analyzed\nto determine the differences in functional groups that appear in FTIR data\nbetween simplicia and 96% ethanol extract. The following are spectra images and\ninterpretation of FTIR data.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 2: FTIR spectra and functional group analysis of shell simplicia of sea urchin.<\/strong><\/p>\n

<\/p>\n

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

Notes : A is the OH group of alcohol, phenol; B is the\nC-H bond of the alkene which strengthens the OH group of the phenol alcohol; C\nis the weak triple C bond of alkyne (usually found in the aromatic group); D is\nan ester with a C-O-H bond that strengthens the OH group of phenol alcohol with\na CH3 bond; E is a C-O bond, this bond exists because in A there is an OH group\nof alcohol, phenol. E strengthens the carboxylic acid; F is the =C-H double\nbond in aromatics.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 3: FTIR spectra and functional group analysis of 96% ethanol shell extracts of sea urchin.<\/strong><\/p>\n

<\/p>\n

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

Notes\n: A is the OH group of alcohol, phenol; B is the alkene C-H bond that\nstrengthens the OH groups of phenol alcohols and amides<\/p>\n\n\n\n

C\nis the aromatic ring; D is an ester with a C-O-H bond that strengthens the OH\ngroup of phenol alcohol; E is an ester with a C-O-H bond that strengthens the\nOH group of phenol alcohol<\/p>\n\n\n\n

The FTIR results\nabove between simplicia and extracts show a slight difference. It appears in\nthe FTIR spectra of the extract. In the simplicia sample, there is no splitting\nprocess known as extraction so the results seen in the spectra generally start\nwith the OH, CH, CO, and CH3 groups. In simplicia, the aromatic and ester ring\nstructures are visible. The extracted sample looks more detailed, and this is\ndue to the extraction process in which the sample is broken down so that the\nspectra results are more detailed, one of which is the presence of N-H, C = C,\nand C-O groups. The details of the types of bonds in the extract could be seen\nwhich show the presence of aromatic alcohols, esters and phenols, in addition\nto the presence of amide groups. The extraction process can break down the\nchemical content of plants or animals. It can be concluded that the extraction\nprocess affects the groups produced by FTIR. The two FTIR spectra results above\nstrengthen the results of phytochemical screening, which are positive for\nflavonoids, tannins, saponins, and alkaloids. Specific groups of these\ncompounds are found in the spectra above. They start from alcohol, aromatic,\nketone, ether, and double bond.<\/p>\n\n\n\n

Results of Antioxidant Test of EEDP\nwith DPPH method<\/strong><\/p>\n\n\n\n

DPPH is one of the methods that\ncan be used in antioxidant testing. The presence of antioxidants in 96% ethanol\nextract of Diadema paucispinum<\/em> sea urchin shells will neutralize the\nDPPH radical by giving electrons to DPPH, resulting in a color change from\npurple to yellow or the intensity of the purple solution reduced.<\/p>\n\n\n\n

The higher the concentration of\nantioxidant chemicals in a sample, the more compounds will donate electrons or\nhydrogen atoms to DPPH free radicals, causing DPPH color fading. When DPPH is\nexposed to substantial concentrations of antioxidant chemicals, it turns from\ndark purple to yellow. This shift in DPPH color is also connected to the energy\nthat DPPH free radicals carry. When in a radical state, DPPH is unstable\n(reactive) and has a high energy because it constantly reacts to find its\nelectron pair, but once found, DPPH becomes more stable (low energy).<\/p>\n\n\n\n

The IC50 value is a number that\nrepresents a 50% reduction in DPPH oxidation (capable of lowering DPPH\noxidation by 50%). Several 0% indicates that there is no antioxidant activity.\nIn comparison, a value of 100% means total attenuation and the test must be\nrepeated by diluting the test solution to determine the activity concentration\nlimit. The results of the calculations are placed into the regression equation\n(Y=AX+B), with the extract concentration (ppm) as the abscissa (X-axis) and the\n% reduction value (antioxidant) as the coordinate (Y-axis). This equation\ncalculates the IC50 of each sample, which is represented by a y value of 50 and\nthe x value obtained as IC50. A chemical is stated to be a powerful antioxidant\nif the IC50 value is less than 50 ppm, a potent antioxidant if the IC50 value\nis 50-100 ppm, a medium antioxidant if the IC50 value is 100-150 ppm, and a\nweak antioxidant if the IC50 value is 150-200 ppm. that the antioxidants in the\nsample reduced because they were easily damaged by the external environment,\nlimiting their activity in reducing DPPH free radicals.<\/p>\n\n\n\n

Table 3: Antioxidant Activity Category<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n
\n

No<\/strong><\/p>\n<\/td>\n

\n

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

\n

IC50 <\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

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

\n

 < 50 ppm<\/p>\n<\/td>\n<\/tr>\n

\n

2<\/p>\n<\/td>\n

\n

Strong<\/p>\n<\/td>\n

\n

50 \u2013 100 ppm<\/p>\n<\/td>\n<\/tr>\n

\n

3<\/p>\n<\/td>\n

\n

Moderate<\/p>\n<\/td>\n

\n

100 \u2013 150 ppm<\/p>\n<\/td>\n<\/tr>\n

\n

4<\/p>\n<\/td>\n

\n

Weak<\/p>\n<\/td>\n

\n

150 \u2013 200 ppm<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Table 4: Percentage of Vitamin C Immersion as Antioxidant by DPPH Method<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

ppm (preparation)<\/strong><\/p>\n<\/td>\n

\n

ppm (test)<\/strong><\/p>\n<\/td>\n

\n

DPPH Abs <\/strong><\/p>\n<\/td>\n

\n

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

\n

% immersion<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

5070<\/p>\n<\/td>\n

\n

1014<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.785<\/p>\n<\/td>\n

\n

8.401400233<\/p>\n<\/td>\n<\/tr>\n

\n

10140<\/p>\n<\/td>\n

\n

2028<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.711<\/p>\n<\/td>\n

\n

17.03617270<\/p>\n<\/td>\n<\/tr>\n

\n

15210<\/p>\n<\/td>\n

\n

3042<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.617<\/p>\n<\/td>\n

\n

28.00466744<\/p>\n<\/td>\n<\/tr>\n

\n

20280<\/p>\n<\/td>\n

\n

4056<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.571<\/p>\n<\/td>\n

\n

33.37222870<\/p>\n<\/td>\n<\/tr>\n

\n

25350<\/p>\n<\/td>\n

\n

5070<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.485<\/p>\n<\/td>\n

\n

43.40723454<\/p>\n<\/td>\n<\/tr>\n

\n

30420<\/p>\n<\/td>\n

\n

6084<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.428<\/p>\n<\/td>\n

\n

50.05834306<\/p>\n<\/td>\n<\/tr>\n

\n

40560<\/p>\n<\/td>\n

\n

8112<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.396<\/p>\n<\/td>\n

\n

53.79229872<\/p>\n<\/td>\n<\/tr>\n

\n

50700<\/p>\n<\/td>\n

\n

10140<\/p>\n<\/td>\n

\n

0.857<\/p>\n<\/td>\n

\n

0.339<\/p>\n<\/td>\n

\n

60.44340723<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 4: Graph of % Vit C Immersion versus Grade.<\/strong><\/p>\n

<\/p>\n

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

Table 5: Percentage of Immersion of Sea Urchin Extract Samples in DPPH<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

ppm (preparation)<\/strong><\/p>\n<\/td>\n

\n

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

\n

% immersion<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

5070<\/p>\n<\/td>\n

\n

1014<\/p>\n<\/td>\n

\n

8.401400233<\/p>\n<\/td>\n<\/tr>\n

\n

10140<\/p>\n<\/td>\n

\n

2028<\/p>\n<\/td>\n

\n

17.03617270<\/p>\n<\/td>\n<\/tr>\n

\n

15210<\/p>\n<\/td>\n

\n

3042<\/p>\n<\/td>\n

\n

28.00466744<\/p>\n<\/td>\n<\/tr>\n

\n

20280<\/p>\n<\/td>\n

\n

4056<\/p>\n<\/td>\n

\n

33.37222870<\/p>\n<\/td>\n<\/tr>\n

\n

25350<\/p>\n<\/td>\n

\n

5070<\/p>\n<\/td>\n

\n

43.40723454<\/p>\n<\/td>\n<\/tr>\n

\n

30420<\/p>\n<\/td>\n

\n

6084<\/p>\n<\/td>\n

\n

50.05834306<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 5: Graph of % Immersion of Sea Urchin Extract Samples versus Levels.<\/strong><\/p>\n

<\/p>\n

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

The results and curves in the two tables above\nexplain that the sea urchin extract sample has a better regression when\ncompared to vitamin C. The value of r close to 1 states that the more linear\nthe data. This regression equation will be able to determine the size of the\nIC50.<\/p>\n\n\n\n

Results of ABTS Method Antioxidant\nTesting of Sea urchin (Diadema\npaucispinum<\/em>) samples<\/strong><\/p>\n\n\n\n

ABTS method antioxidant activity\ntesting of sea urchin (Diadema paucispinum) samples is based on the ability of\nantioxidant compounds to stabilize free radical compounds by donating proton\nradicals. The ability of sea urchin extract (Diadema paucispinum) to stabilize\nfree radical compounds can be seen from the change in color of the blue-green\ntest solution to colorless or reduced color intensity. The results of\nantioxidant activity of sea urchin extract (Diadema paucispinum) and vitamin C\nas a comparison. The data shown in the table below.<\/p>\n\n\n\n

Compared to other procedures, the\nABTS method reacts rapidly, and the test is straightforward to repeat. A high\nIC50 value suggests that the antioxidant activity is strong. The IC50 value in\nthe sample solution is the concentration required to lower 50% of the ABTS free\nradical activity. The lower the content in the sample, the lower the absorbance\nvalue and the higher the antioxidant activity.<\/p>\n\n\n\n

Table 6: Percentage of Vitamin C Immersion as Antioxidant by ABTS Method<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

ppm (prepartion)<\/strong><\/p>\n<\/td>\n

\n

ppm (test)<\/strong><\/p>\n<\/td>\n

\n

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

\n

Abs (734 nm)<\/strong><\/p>\n<\/td>\n

\n

% immersion<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

  5070<\/p>\n<\/td>\n

\n

253.5<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.519<\/p>\n<\/td>\n

\n

13.06532663<\/p>\n<\/td>\n<\/tr>\n

\n

10140<\/p>\n<\/td>\n

\n

507.0<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.366<\/p>\n<\/td>\n

\n

38.69346734<\/p>\n<\/td>\n<\/tr>\n

\n

15210<\/p>\n<\/td>\n

\n

760.5<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.283<\/p>\n<\/td>\n

\n

52.59631491<\/p>\n<\/td>\n<\/tr>\n

\n

20280<\/p>\n<\/td>\n

\n

1014.0<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.211<\/p>\n<\/td>\n

\n

64.65661642<\/p>\n<\/td>\n<\/tr>\n

\n

25350<\/p>\n<\/td>\n

\n

1267.5<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.186<\/p>\n<\/td>\n

\n

68.84422111<\/p>\n<\/td>\n<\/tr>\n

\n

30420<\/p>\n<\/td>\n

\n

1521.0<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.161<\/p>\n<\/td>\n

\n

73.03182580<\/p>\n<\/td>\n<\/tr>\n

\n

40560<\/p>\n<\/td>\n

\n

2028.0<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.194<\/p>\n<\/td>\n

\n

67.50418760<\/p>\n<\/td>\n<\/tr>\n

\n

50700<\/p>\n<\/td>\n

\n

2535.0<\/p>\n<\/td>\n

\n

0.597<\/p>\n<\/td>\n

\n

0.242<\/p>\n<\/td>\n

\n

59.46398660<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 6: Graph of % Immersion of Vit C in ABTS Reagent versus Grade.<\/strong><\/p>\n

<\/p>\n

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

Table 7: Percentage of Immersion of samples of Sea urchin Extract as Antioxidant by ABTS Method<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n\n
\n

ppm (preparation)<\/strong><\/p>\n<\/td>\n

\n

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

\n

% immersion<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

5070<\/p>\n<\/td>\n

\n

253.5<\/p>\n<\/td>\n

\n

13.0653266<\/p>\n<\/td>\n<\/tr>\n

\n

10140<\/p>\n<\/td>\n

\n

507<\/p>\n<\/td>\n

\n

38.6934673<\/p>\n<\/td>\n<\/tr>\n

\n

15210<\/p>\n<\/td>\n

\n

760.5<\/p>\n<\/td>\n

\n

52.5963149<\/p>\n<\/td>\n<\/tr>\n

\n

20280<\/p>\n<\/td>\n

\n

1014<\/p>\n<\/td>\n

\n

64.6566164<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 7: Graph of % Immersion of Sea urchin Samples in ABTS Reagent versus Levels<\/strong><\/p>\n

<\/p>\n

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

Table 8: Antioxidant Activity of Vitamin C and Diadema paucispinum<\/em> Extracts<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

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

\n

IC50<\/sub> <\/strong>DPPH method<\/strong><\/p>\n<\/td>\n

\n

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

\n

Vitamin C<\/p>\n<\/td>\n

\n

3,25 ppm<\/p>\n<\/td>\n

\n

2,09 ppm<\/p>\n<\/td>\n<\/tr>\n

\n

Diadema paucispinum<\/em> extract<\/p>\n<\/td>\n

\n

6084 ppm<\/p>\n<\/td>\n

\n

756,3 ppm<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

In determining the\nantioxidant activity, the IC50 parameter is used, which is the sample\nconcentration required to capture DPPH radicals by 50%, where the smaller the\nIC50 value, the stronger the antioxidant activity. Meanwhile, Molyneux (2004)\ncategorizes an IC50 value below 50 ppm as a powerful antioxidant, an IC50 value\nof 50 – 100 ppm as a potent antioxidant, an IC50 value of 100 – 150 ppm as a\nmedium antioxidant, and an IC50 value of 150 – 200 ppm as a weak antioxidant.\nBased on the results of antioxidant activity testing of sea urchin extract (Diadema\npaucispinum)<\/em> with DPPH and ABTS immersion methods, the extract did not have\nantioxidant properties because the IC50 value was > 200 ppm. In contrast,\nVitamin C as a standard drug had a solid value as an antioxidant.. <\/p>\n\n\n\n

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

Based on\nthe results of the study, it was found that 96% ethanol extract of Diadema\npauscispinum<\/em> sea urchin found in Sumenep-Madura contains alkaloids,\nflavonoids, saponins, and tannins. Antioxidant activity of 96% ethanol extract\nof Diadema pauscispinum<\/em> sea urchin showed with IC50 value greater than\nvitamin C, this indicates that 96% ethanol extract of Diadema pauscispinum<\/em>\nsea urchin did not have antioxidant activity compared with vitamin C as a\nstandard compound.<\/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 are no\nconflicts of interest.<\/p>\n\n\n\n

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

This project was funded by Ministry of Research and Technology for Higher Education (Kemendikbudristek DIKTI) contract number 032\/SP2H\/PT\/LL7\/2023; B\/13\/HIB-EX.PB\/UHT.C7\/VI\/2023. <\/p>\n\n\n\n

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

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    Introduction Sea urchin, locally known as sea urchin\/tehe-tehe, is a […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[111],"tags":[],"class_list":["post-54427","post","type-post","status-publish","format-standard","hentry","category-vol16no4"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/54427","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=54427"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/54427\/revisions"}],"predecessor-version":[{"id":55186,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/54427\/revisions\/55186"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=54427"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=54427"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=54427"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}