RJOAS, 5(77), May 2018

DOI https://doi.org/10.18551/rjoas.2018-05.38

EVALUATION OF CHLOROPHYLL, TANIN, GROSS ENERGY AND THE WOMB NUTRIENTS ON THE LEAVES AND THE SEEDS OF INDIGOFERA SP.

Atma Alfian Adi Graduate Student, Faculty of Animal Husbandry ,

Sjofjan Osfar, Djunaidi Irfan H. Lecturers of Nutrition and Animal Feed Division, Faculty of Animal Husbandry, University of Brawijaya, Indonesia

*E-mail: [email protected]

ABSTRACT This research aims to evaluate the seeds and leaves of with the optimal level to the addition of material feeds in the form of Indigofera sp. flour. The result showed that the womb of chlorophyll of Indigofera sp. for its seeds were 159.5p glg, while for its leaves were 339,3p glg. Tanin’s identification of Indigofera sp. for its seeds were 0,84 ± 0,02 and for its leaves were 1,47 ± 0,01. The results of gross energy on Indigofera sp. were 4,421 for its seeds and 4,229 for its leaves.

KEY WORDS Chlorophyll, tanin, gross energy, proximate analysis, Indigofera sp.

Indigofera sp plants have high productivity and the womb nutrient was good enough, especially the high protein of its womb. Tarigan et al. (2010) stated that the production of dry matter of Indigofera sp plant which was cut at 60 days old with 1.0 m cutting height was 31.2 ton / ha / year, which was the highest production when it compared to the age of the older cutting or the younger ones. Then, at 60 days of cutting age, higher crude protein content was found to be higher if it compared to 90 days or 30 days. This plant was also very tolerant for dry season and puddles that can be planted in the territory of Indonesia. This plant can be used as animal feed rich in nitrogen, phosphorus and calcium. Indigofera sp. was very well used as forages of animal feed and contains crude protein 27.9%, crude fiber 15.25%, 0.22% calcium and 0.18% phosphorus. Legum Indigofera sp. has high protein content, tolerance to drought, waterlogging and salinity tolerance (Hassen et al., 2007). Tarigan (2010) mentioned that the content of crude protein, calcium, and phosphorus decreased with increasing cut interval, while the organic matter content, NDF, ADF was higher with increasing cut interval. The decrement in the womb of nutrient with increasing age of plants can be illustrated by the ratio of leaves / stems to plant. Leav’se blade have higher womb of protein when its compared to the stem part of the plant (Whitehead 2000). This also means that the protein content of the stems tends to be lower than that of the leafy fraction protein. White et al. (2001) suggested that yields were more responsive for spacing compared to the plant populations. Shortening the spacing from 100-70 cm will increase the production of Navy Bean by 17%. Akbarillah et al. (2002) reported that nutritional value of Indigofera sp flour as follows: crude protein 27.97%; crude fiber 15.25%. Indigofera sp spoons were a source of protein and contain high pigments such as xantophils and carotenoids. In vivo digestibility test of the potential of Indigofera sp leaves as Boerka goat feed has been investigated by Tarigan (2010). Indigofera sp feed given the result of interval treatment and the best cutting intensity that was cutting interval 60 days and cutting height of 1.5 meter. Based on the above background, this research was conducted to evaluate the best womb nutrient between the seeds and the leaves of Indigofera sp.

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MATERIALS AND METHODS OF RESEARCH

Laboratory Service Unit MRCPP Ma Chung University , Clinical Pathology Laboratory Faculty of Medicine Universitas Brawijaya Malang, Chemistry Laboratory Universitas Brawijaya Faculty of Mathematics and Natural Sciences. Leaves Flour Production Procedure Indigofera sp. The materials used in the study were Indigofera sp leaves harvested at 60 days. Indigofera sp. beginning with the harvesting of Indigofera sp plants that have 60 days defoliation age, by cutting the shoots <0.5 cm in diameter. Then followed by drying the leaves of Indigofera sp in greenhouse to dry in order not to cause green color change. It was further ground into flour, and ready for use in the preparation of feed. The Procedure of Making Indigofera sp. The material used in the research was the seeds of Indigofera sp which have been harvested in the drying and then peeled from the skin, then milled to become flour and ready for use in the preparation of feed. Procedure Chlorophyll Level. The procedure for determining chlorophyll concentration is done by the International Rice Research Institute (IRRI) method modified by Balitbio Bogor (Alsuhendra, 2004). A total of 0.1 g of green cincau leaves, pegagan leaf, katuk leaf, mulberry leaf, which has been chopped (about 2 mm in size), put into a test tube, then 20 ml of acetone added. The mixture is shaken sufficiently, and then stand for 2 × 24 hours. The extract obtained was analyzed by concentration of chlorophyll a and chlorophyll b using spectrophotometer, each at 645 and 663 nm wavelengths. Procedures Determination of Total Tannin. The procedure of determining the womb of tannin was done with, as much as 50.0 mg of ethanol extract 70% Leaf Indigofera sp dissolved with aqua demineralisata until volume 50,0 ml. The extract solution obtained was then dipipet a certain amount and added 1 ml of Folin Ciocalteu reagent, then shaken and allowed to stand for 5 minutes. To the solution plus 2 ml of 15% Na 2 CO 3 solution, shake homogeneously and stand for 5 minutes. Furthermore, aqua demineralisata added to volume 10.0 ml, let stand in stable time range obtained. Absorbance of the extract solution was observed at maximum wavelength. The concentration obtained was replicated twice. Total tannin content was calculated equivalent to galic acid (Gallic Acid Equivalent / GAE). Gross Energy Determination Procedure (GE). The procedure of determining gross energy by using the Bomb Calorimeter method, principally by the amount of heat measured in calories, produced when a material or sample was completely oxidized in a calorimeter bomb as called the total energy of the material or sample. In the determination of this total energy occurs the conversion of chemical energy in a material or sample.

RESULTS AND DISCUSSION

Chlorophyll levels. Chlorophyll was a pigment compound that plays a role in selecting the wavelength of light that energy is being taken in photosynthesis. Actually, the leaves also have other pigments, such as carotene (colored orange), xantofil (yellow), and anthocyanin (red, blue, or purple, depending on the degree of acidity) (Anonymous 2006). All types of plastids including chlorophyll were derived from protoplastids, a colorless organelle found in plant cells that grow in dark and light (Lakitan 2001). The synthesis of chlorophyll a from chlorophyll a does not require light, The change of protoklorofilid to chlorophyll a in Angiospermae absolutely requires light, but on Gymnospermae (some ferns and algae, chlorophyll can be formed in dark circumstances. The degree of the leaves development was the synthesis of chlorophyll b from chlorophyll a with a large number, followed by the development of the leaf. Chlorophyll synthesis b continues along with The development of leaves characterized by the change of green leaf color to dark green. The womb of Chlorophyll in dark green leaf was 72% larger than the leaves of light green color. Chlorophyll b was formed from chlorophyll a or chlorophyll a Chlorophyll a and b are the main pigments contained in the thylakoid membrane. In addition to these two

321 RJOAS, 5(77), May 2018 pigments there were also yellow to orange pigments called carotenoids. There were two types of carotenoids: carotene (pure hydrocarbons) and xantofil (containing oxygen) (Lakitan 2001). This research uses HPLC method where to determine the level of Chlorophyll on Indigofera sp plant the sample used in this research is the seeds and leaves of Indigofera sp. Based on Table of Chlorophyll content on Indigofera sp plant seeds 159.5p glg while on leaf 339,3p glg and based on the analysis on leaf Chlorophyll content is highest compared with Indigofera sp seeds (Table 1). Factors that affect chlorophyll formation include gene, light (Subandi 2008).

Table 1 – Sometimes Chlorophyll Leaves and Seeds of plants Indigofera sp.

Plant Leaf Seed Indigofera sp 339,3p glg 159,5p glg

Source: MRCPP MACHUNG Malang Laboratory.

Tanin levels. Tanin was a very complex organic substance and consists of many phenolic compounds found in various plants. Generally, tannin was scattered almost in all parts of the plant as in the bark, stems, leaves, seeds and fruit (Sajaratud, 2013). Tanin is an active compound of secondary metabolites that are known to have several properties such as astringent, anti diarrhea, antibacterial and antioxidants (Desmiaty et al., 2008). The results of the experimental species of tannins of Indigofera sp on seeds and leaves can be seen in Table 2. The result of identification of tannin species of Indigofera sp on seeds 0.84 ± 0.02 and leaf 1.47 ± 0.01 based on the analysis on Indigofera sp plant tannin content on leaves was higher than the seeds. This was comparable to previous research has been done determination of tannins and total tannin content on the leaves and seeds of Indigofera sp plant by colorimetry (Desmiaty et al., 2008) and the determination of the womb of tannin on the leaves of Indigofera sp by permanganometri and kolorimetri. This research wass used the part of the leaves from Indigofera sp plants that were old to see the levels of tannin compounds because the leaves were still old containing compounds of tannins were relatively higher.

Table 2 – Tanin Level on Indigofera sp.

Plant Leaf Seed Indigofera sp 1,47 ± 0,01 0,84 ± 0,02

Source: UPT Analytical Service and Measurement Department of Chemistry FMIPA Universitas Brawijaya Malang.

Gross energy. The energy needs of livestock met from food, therefore the first step that needs to be considered in preparing the ration was to provide for the needs of the fodder substances, and the second one was to choose the food that will be used to meet those needs. One example was the feed material from Indigofera sp. The stages in this study distinguish between seeds and leaves of Indigofera sp plant which can be seen in table 3. The results showed that GE (Gross Energy) on Indigofera sp. were 4,421 and 4,229 for its leaves. Many factors affect the results of livestock research or nutrients from plants. Among them depends on the type, age on the plant. Energy determination total samples to be analyzed were usually in solid form and comprise organic and inorganic compounds. In the event of combustion of the material, the organic compounds are usually composed of elements C, H, 0, N, P and S.

Table 3 – Gross Energy Analysis of Plants Indigofera sp.

Plant Leaf Seed Indigofera sp 4.229 4.421

Source: Chemistry Laboratory State University of Malang (UM).

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Food Analysis based on Proximate Analysis. The results of the study based on the proximate analysis on Indigofera sp sample samples that distinguish parts of the seeds and leaves were shown in Table 4. The quality of a plant to be used as a feed can be determined by looking at the content of food substances in the plant. The content of a plant's food substances can be obtained through proximate analysis methods in the laboratory

Table 4 – Content of Plant Proximate Analysis Indigofera sp.

Feed ingredients Dry matter (%) Ash (%) Crude protein (%) Coarse fiber (%) Rough fat (%) Leaves of Indigofera sp 93,30 12,16 25,77 13,45 2,52 Seeds of Indigofera sp 91,55 5,51 28,50 16,71 1,41

Source: Result of Laboratory Analysis of NMT (2018) Faculty of Animal Husbandry Universitas Brawijaya Malang.

Dry Material. Based on the variance analysis in table 4, the dry matter proximate test showed that the Indigofera sp on the seeds showed 91.55 and the leaves showed 93.30. The high content of dry ingredients affects the low water content in the feed. The high moisture content of young plants because in the young plants there are more active cells when compared to old plants that have more cells damaged or dead in it. The water itself in the leaf cell is required as a substrate or reactant for various biochemical reactions such as photosynthesis. In addition, water also serves as a nutrient solvent so that it can transport nutrients to the leaves (ishaq, 2001). Ash Content. Result of Proximate Analysis of ash content in table 4, Indigofera sp plant showed that on leaf 12,16 and at seed 5,51. The high ash content on the leaf plant because the plant part of the leaf there are more active cells when compared to seed plants that have more cells damaged or die in it, so that the nutrients absorbed by the roots were all brought to the leaves. Ash content on the part of the seeds were not go up because of the seeds in the death of cells. The death of these cells causes the nutrients absorbed by the roots that can not be maximally brought to the seeds (ishaq, 2001). Crude protein. Raw protein analysis results in Table 4, proximate test of Indigofera sp showed that the leaves were 25.77 and the seeds were 28.50. The high levels of plant protein part of the seeds were caused by the function of proteins used as the forming of cells, tissues, and plant organs as well as functioning as synthetic materials of chlorophyll, enzymes, and amino acids are more common in seed plants than leaves (ishaq, 2001). Coarse Fiber. Result of analysis of crude fiber in table 4, proximate test of Indigofera sp showed that leaves were 13.45 and the seeds were 16,71. The high levels of crude fiber in the seeds happen because the content of coarse fiber was closely related to the age of the plant. The older the age of the plant was increasing the content of crude fiber, especially in the seed plant (ishaq, 2001). Rough Fat. Result of analysis of crude fat in table 4, proximate test of Indigofera sp leaf part 2,52 and seed part 1.41. Classification of fats and oils According to Harris et al (2000), based on its structure the fat consists of simple lipids Fatty alcohol esters for example: glyceride, fat, and night esters; composite lipids and sphingolipids Non-alcohol fatty esters for example: phospholipids, glycolipids, aminolipids, lipoproteins; derived lipids for example: fatty acids, glycerol, ketones, hormones, fat soluble vitamins, steroids, carotenoids, fatty acid aldehydes, waxes and hydrocarbons.

CONCLUSION

Based on the results of this research on Indigofera sp plant, between seeds and leaves showed that Indigofera sp flour was the best plant because it has the high womb of chlorophyll, and also the chlorophyll can aim to affect egg quality, and Indigofera sp leaves have high content of Tanin, BK, Abu, and LK levels compared to seeds Indigofera sp, but leaves Indigofera sp have low GE, PK, SK content compared to seeds Indigofera sp.

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