INTRODUCTION:

Flavonoids are the pigments present in plants which mostly found in terrestrial plants. Flavonoids are indeed a naturally present group of polyphenolic compounds present in plants¹. The greatest category of natural phenolics is the flavonoid, and both woody and non-woody plants have been shown to contain more than 2,000 flavonoids².

Plants produce a wide variety of secondary metabolites that they use to interact with their surroundings. The medicinal effects of several of these substances as well as their industrial or agricultural uses raise the crops' market worth³.The traditional medicine all over the world now a days is revealed by an extensive activity of research on different plant species and their therapeutic principles⁴. As plants produce a lot of antioxidants to control the oxidative stress caused by sun beams and oxygen, they can represent a source of new compounds with antioxidant activity⁵. Free radicals are produced as a part of normal metabolic processes⁶. Antioxidants obtained from traditional medicinal plants may therefore offer protection against a range of illnesses and slow down oxidation processes in food systems⁷.

Red algae (Eucheuma denticulatum (Burman) Collins et Harvey) are environmentally considered to be the major providers and structural producers of habitat for many other marine organisms, and their important role is in the primary integration and protection, maintenance and maintenance of coral reefs⁸. The presence of different bioactive molecules in seaweed is thought to be due to environmental conditions and the habitat of these marine organisms in natural aquatic communities, where there is an inhibitory competition between producers and consumers leading to chemical weapons enslavement with very high targets and strong effects when diluted with lots of creases with water⁹.

Red algae (Rhodophyceae) occupy the highest number of species that grow in Indonesian sea waters, namely around 452 species, after that green algae (Chlorophyceae) are around 196 species and brown algae (Phaeophyceae) are around 134¹⁰.Behind its ecological and biological role in maintaining the stability of marine ecosystems and as a place to live as well as protection for other biota, this macroalgae group has economic potential, namely as a raw material for industry and health^11,12.

Tempeh is generally used as a vegetarian protein source. Driven by its high nutritional value, too, much research has focused on the use of tempeh and tempeh-like products in relation to malnutrition in Third World countries¹³. Products such as tempeh are produced from legumes other than soybeans as well as cereals (eg wheat or corn), and cereal/soy mixtures are used as starting materials for fermentation. For economic reasons in Indonesia, more and more tempeh-like products are circulating in the market. In particular, products such as tempeh made from corn/soybens (30:70) are produced, as the cost for corn is lower than for soybeans¹⁴. Tempeh is a fermented food made from soybeans and is a nutritious, affordable and sustainable source of functional protein. Globally, tempeh is a fermented product that is widely accepted. Although there is a lot of literature on tempeh, most of the research focuses on unfermented soybeans, so the impact of tempeh fermentation on the biological properties of soybeans is still largely scattered^15,16,17.

Numerous natural and synthetic chemicals, including phenolic resins, herbicides, surfactants, alkaloids, steroids, and glycosides, are members of the phenolic group¹⁸. Secondary metabolites known as phenolic compounds are produced by plants¹⁹. Polyphenolic antioxidants can obstruct the oxidation process through a variety of mechanisms, such as the compound's reducing capability²⁰.

MATERIALS AND METHODS:

Preparation of Red Algae:

Red algae were obtained from Bantaeng district in Indonesia. Then determination of red algae species was carried out at the department of biology, Hasanuddin University, Indonesia.

Process of Making Tempeh:²¹

Soybeans are washed under running water until clean, then soak the soybeans for 5 hours.When finished soaking, then wash the soybean soaking again until clean.Next, boil the soaked soybeans that have been washed clean for 30-45 minutes, then soak overnight.The next step is to peel the epidermis of the soybeans that have been soaked overnight by hand or using a machine, then wash them thoroughly.Steam for 20 minutes, then remove and cool for a while then sprinkle with tempeh yeast and stir until blended.After that, wrap the soybeans using banana leaves or plastic, as desired.The fermentation process will also take 2 days at normal room temperature. If the soybeans have been covered by the fungus evenly, then the tempe is cooked and ready to be processed.

Extraction of Red Algae (Eucheuma denticulatum (Burman) Collins et Harvey) Using Kinetic Maceration Method:²²

3.74 kg of dry Red Algae samples dissolved in water with a ratio of 1: 2 for 2 repetitions with a temperature of 500 C while stirring for about 1 hour. Then the results of the extraction were filtered using a filter and the results of the extraction were stored in the freezer with a temperature of -300C for 50 hours, then put into Freeze Dry for 24 hours. The yield of the extract was then calculated by comparing the initial weight of the simplicia with the final weight method of the resulting extract.

Extraction of Tempeh Using Kinetic Maceration Method:²²

5.145 kg of Tempe samples dissolved in water with a ratio of 1: 2 for 2 repetitions with a temperature of 500 C while stirring for about 1 hour. Then the results of the extraction were filtered using a filter and the results of the extraction were stored in the freezer with a temperature of -300C for 50 hours, then put into Freeze Dry for 24 hours. The yield of the extract was then calculated by comparing the initial weight of the simplicia with the final weight method of the resulting extract.

Analysis of Total Flavonoid (Castejon):²³.

Weigh 1 gram of sample and add 10 ml of distilled water, Leave it overnight, Pipette approximately 2 ml of solution and react with 100 micron AlCl₃ 10% and 100 micron sodium acetate 1 M. After that incubation for 30 minutes. Measure the absorbance at a wavelength of 430 nm, create a standard curve using quercetin.

Analysis Of Total Phenolic^24,25.

Weigh the sample 0.5-1 gr, add 10 ml of 96% alcohol and leave overnight. Pipette 0.5 ml of sample then add 0.5 ml of folin then stir and leave for 5 minutes, add 0.5 ml of saturated sodium carbonate, add 3.5 ml of distilled water and then shake. Leave it for 60 minutes in a dark room. Centrifuge and measure on a spectrophotometer with a wavelength of 725nm, create a standard curve using different concentrations of gallic acid.

RESULTS:

Table 1: Results of Testing for total Flavonoid and Phenolic Content of Red Algae Extract

Sample	Flavonoid (mg/kg)	Phenolic (mg/kg)
A Red Alga Extract	54.26	400.45
B Red Alga Extract	54.57	432.62
C Red Alga Extract	56.94	428.61
Mean	55.25	420.56

Table 2: Absorbance of The Quercetin Calibration Curve of Red Algae

Concentration (mg/l)	Absorbance
0	0
2	0.196
4	0.365
6	0.525
8	0.709
10	0.839

Figure 1: Flavonoid Curve of Red algae Extract

Table 3 : Absorbance of The Gallic Acid Calibration Curve of Red Algae

Concentration (mg/l)	Absorbance
0	0
10	0.289
20	0.443
40	0.713
80	1.262
160	2.300

Figure 2 . Polifenol Curve of red algae Extract

Figure 3. Flavonoid Curve of Tempeh Extract

Table 6: Absorbance of The Gallic Acid Calibration Curve of Tempeh Extract

Concentration (mg/l)	Absorbance
0	0
10	0.289
20	0.443
40	0.713
80	1.262
160	2.300

Figure 4: Polifenol Curve of TempehExtract

DISCUSSION:

Antioxidant compounds are oneone of the most sought-after compoundsresearcher. Antioxidant compounds cancounteract free radicals, which are causedby pollution. Free radicals canenter through inhalation or foodinto the human body. Free radicalsare atoms or moleculesloses an electron pair on the surfaceouter skin. Free radicals are easy to react withmolecules such as proteins, fats,carbohydrates, and DNA. Free radicals cancause various diseaseswhich can be prevented by compoundsnatural antioxidants.

Flavonoids are the largest group of phenolic compounds that have chromophore groups. This group has the ability to absorb UV light waves.

Red algae haveantioxidant activity is due tohave the pigments chlorophyll a, chlorophyll d, zeaxanthin, cryptoxanthin lycopene, α-carotene, β-carotene, lutein, and picobilin. Pigmentcarotenoids found in red algaethis is the basic ingredient information of natural antioxidant compounds.

Tempeh is a fermented soybean dish that is well-known for having a lot of protein. In the world, there are various ways to make tempeh, including soaking it in vinegar before fermentation. Water-facilitated extraction, alcalase, and papain hydrolysis, along with membrane filtration, were three preparative techniques used to produce tempeh-based peptides.

CONCLUSIONS:

Red algae and tempeh originating from Indonesia have high levels of flavonoids and phenolics, and are very useful as food ingredients or as a mixture of medicinal ingredients. Total Flavonoid content test for total phenolic content was carried out quantitatively using a uv-vis spectrophotometer at a wavelength of 370 nm. The results showed that the average total flavonoid content in red algae was 55.25 mg/kg, and the total phenolic content in red algae was 420.56 mg/kg. whereas in tempeh the average total flavonoid content in red algae was 223.20 mg/kg, and total phenolic in tempe was 736.40 mg/kg.

CONFLICT OF INTEREST:

The authors have no conflicts of interest regarding this investigation.

ACKNOWLEDGMENTS:

The researcher expresses her highest gratitude to the Indonesia Endowment Fund For Education (LPDP) under the ministry of finance of The Republic Of Indonesia as a sponsor in conducting research and publishing the results of this research.

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Received on 10.05.2023 Modified on 11.07.2023

Research J. Pharm. and Tech 2024; 17(4):1692-1696.

DOI: 10.52711/0974-360X.2024.00268