ABSTRACT This study was conducted to determine the proximate composition and energy value of kek batik.

Standard methods of the Association of Official Analytical Chemists (AOAC) were used to determine the moisture, ash, crude protein, crude fat, and crude fiber contents of the sample. The carbohydrate content in the sample was determined by difference and the energy value was calculated. There are approximately 6.413%, 1.712%, 4.94%, 22.98%, 1.995%, and 61.96% for the moisture, ash, crude protein, crude fat, crude fiber, and carbohydrate contents respectively in kek batik sample. The total energy of our sample calculated is 474.4 kcal.

INTRODUCTION Proximate analysis is analysis of foods and feeding stuffs for nitrogen (for protein), ether extract (for fat), crude fibre and ash (mineral salts), together with soluble carbohydrate calculated by subtracting these values from the total (carbohydrate by difference). The food sample tested in this experiment is batik cake. Batik cake is made up of Marie biscuits, Milo, condensed milk, and butter. It is usually served as dessert. Batik cake is really easy to make. In a pan, place butter and heat over slow fire. Add the condensed milk and Milo to the butter, and stir continuously. Once thickened, add the broken Marie biscuits and turn the heat off. Slowly combine all the biscuits together. Finally, place the mixture in a container. Foods are heterogeneous materials that contain different proportions of chemically bound, physically bound, capillary, trapped or bulk water. Water is the main constituent of most food products and the water content of foods varies widely. Determination of moisture content is of the most important and most widely used measurement in food processing because of the significance effect of water on stability and quality of foods. Water in a food item can be present in 3 ways that is free water, absorbed water and free water. Free water is physically linked to the food matrix and easily lost by evaporation or drying as a separate constituent. Meanwhile, bound water includes water molecules chemically bonded to ionic and polar groups or water of crystallization or hydrates which is difficult to remove. Moisture content is one of the most commonly measured properties of food materials. The texture, taste, appearance and stability of foods depend on the amount of water or moisture they contain.

There are few methods could be applied to determine the moisture content for examples evaporation, distillation, chemical, physical and spectroscopic methods. In this experiment, moisture content of food sample is determined by using oven drying method. Oven drying methods involve the removal of moisture from the sample and the weight loss from sample is used to calculate the moisture content of the sample (Nielsen, 1998). The principle of oven drying method is dry a known weight of food sample in an oven at C and the loss of

weight is used to calculate the moisture content of the sample. The sample is weighed and heated in an oven to constant weight. Ash is the inorganic residue which is remained after burning process of organic compounds in food at temperature 500 to 700 C. Amount and composition of ash depends on methods of ashing for respective foods. Ash content can be used to identify any authenticity in food and also act as basic food component.

There are three methods in determination of ash i.e. dry, wet and conductometric methods. Dry method is normally used for all kinds of food but not suitable for trace metals such as arsenic, mercury or lead. All non-water soluble, water soluble and non-acid soluble ash can be determined using this method. Meanwhile, wet ashing method is more suitable to be used to obtain ash which will be used to determine trace metals and metallic poison. Conductometricis normally used for sugar and high sugar products such as syrup, honey and drinks. Protein are polymers of amino acids the majority of which are -amino acid having the general formula NH2CHRCOOH, and may thus be distinguished from fats and carbohydrate in being the only macronutrient in foods to contain nitrogen. The presence of nitrogen in proteins is often used as the basis of the estimation of proteins in foods. Protein content can be determined with the analyses of any protein component like carbon or nitrogen, amino acid group or peptide chains. Few methods are available for the determination of crude protein but the analyses using nitrogen (N) content are the most prominent and widely used in Kjehdahl method.

The Kjeldahl method is a means of determining the nitrogen content of organic and inorganic substances. Although the technique and apparatus have been altered considerably over the past 100 years, the basic principles introduced by Johan Kjeldahl endure today. The

Kjeldahl method may be broken down into three main steps which are digestion, distillation and titration. Digestion - the decomposition of nitrogen in organic samples utilizing a concentrated acid solution. This is accomplished by boiling a homogeneous sample in concentrated sulfuric acid. The end result is an ammonium sulfate solution. Distillation adding excess base to the acid digestion mixture to convert NH4
+

to NH3, followed by

boiling and condensation of the NH3 gas in a receiving solution. Titration - to quantify the amount of ammonia in the receiving solution. The amount of nitrogen in a sample can be calculated from the quantified amount of ammonia ions in the receiving solution. Fats consist of a wide group of compounds that are generally soluble in organic solvents and largely insoluble in water. Fats may be either solid or liquid at room temperature, depending on their structure and composition. Some fats such as triacylglycerides are very hydrophobic but di- and mono-acyglycerols have both hydrophobic and hydrophilic moieties in their molecules and are soluble in relatively polar solvent. Fats are generally solid at room temperature while oils are liquid at room temperature. Their compositions are generally similar. The determination of fat content in food in this experiment was study to understand the importance of fat in food system and analysis. Instead of fat provide energy, it also allows the absorption of important nutrients, give the sensory characteristic and value added to the food product. The Soxhlet method is used in the analysis, instead of using other method; Gerber, Babcock, Bligh-Dyer, Werner-Schmidt and Rose-Gottlieb. Soxhlet method is widely used in food commodity especially for solid food. In this method, fat is extracted, semicontinuously with the organic solvent. The solvent is heated, volatilized, and condensed above the sample. Fat content of the sample is determined by measuring difference in oven dried sample weight before and after ether extraction. The weight loss of sample is interpreted as fat content. Fat dissolves into the ether which is then squeezed out of the sample.

Fibre is the edible parts of plants or analogous carbohydrates that are resistant to digestion and absorption in the human small intestine with complete or partial fermentation in the large intestine. While crude fibre is the loss on ignition of the dried residue remaining after digestion of the sample with 1.25% sulfuric acid and 1.25% sodium hydroxide solutions under specific conditions (AACC International Method, 1999). Crude fibre is defined as an

organic food residue which has been hydrolyzed by acid and aqueous alkaline. The method for determining crude fibre consists basically of measuring the residue remaining after acid and basic hydrolysis. Crude fibre method measures fibre like cellulose and lignin, therefore there is no specific component being determined. These residues (containing cellulose, hemicellulose, lignin, ash and tannins) are indigestible substances and have low or no nutritional value to us. It is known that the crude fibre method covers 50-80 per cent of the cellulose, 10 - 50 per cent of the lignin, and 20 per cent of the hemicelluloses (J.B. Robertsons 98 ). This method is one of the official AOCS (American Oil Chemists

Society) methods that were used where it is more suitable for oilseeds and high fat foods. In a food, crude fibre is only one-seventh to one-half of total dietary fibre, since not all types of fibre can be determined using crude fibre method (Anderson et al., 2007). In food composition tables, the carbohydrates content has been usually given as total carbohydrates by difference, i.e. the percentage of water, protein, fat and ash subtracted from 100. Another widely used term is nitrogen-free extract, calculated as components other than water, nitrogenous compounds, crude fibre, crude fat and minerals. The increasing awareness that specific carbohydrates play significant metabolic and functional roles and the availability of analytical tools to determine individual components aroused interest in investigations on their distribution in many foods. Carbohydrates determination can be done through qualitative and quantitative analysis. Qualitative analysis like colour tests, paper chromatography, ensures that ingredient labels present accurate compositional information. Whereas quantitative analysis ensures added components are listed in the proper order on ingredient labels. MATERIALS Determination of Moisture Content Materials used in this study are crucible, drying oven at 101-1050C, analytical balance (0.1 mg sensitivity), glass rod, desiccators, tongs, spatula and batik cake as the sample Determination of Ash Content Materials used in this study are sample (batik cake), muffle furnace at 550C, crucible, thong, desiccator and spatula Determination of Crude Protein

Materials used in this study are concentrated sulphuric acid (nitrogen-free), mix catalyst (96% sodium sulphate anhydrous + 3.5% cuprum sulphate + 0.5% selenium dioxide), sodium hydroxide solution : 45% (w/v), distilled water, indicator solution : 0.2% methyl red + 0.1% methylene blue in 96% ethanol, 0.5N HCl (for reverse titration) or 2% boric acid (for direct titration), 0.5N NaOH (for reverse titration) or 0.05N H2SO4 (for direct titration) and batik cake.

Determination of Crude Fat In this experiment, we used batik cake too as our sample product. We also used petroleum ether as solvent. The apparatus needed for this experiment is Soxhlet apparatus, round bottom flask, cotton wool, thimble, beaker, measuring cylinder, spatula and desiccators. Determination of Crude Fibre Materials used in this study are sulphuric acid: 0.25N, sodium hydroxide: 0.313N, alcohol and sample (batik cake), muffle furnace set at 550C, Buchner funnel or glass funnel, spatula, heater, conical flask, desiccator, oven set at 105C, crucible, filter paper, condenser, ashless filter paper No. 541 and aspirator pump.

PROCEDURE

Determination of Moisture Content First of all, the oven was heated to 105oc and the temperature was kept constant. After that , all three crucibles and its covers were cleaned and dried in the oven for a minimum 30 minutes. After 30 minutes, the crucibles and its covers were transferred into desiccators by using a tongs and let the crucibles cool in the desiccators for approximately 20 minutes. Then, weight all three crucibles and its covers rapidly and accurately by using analytical balance. The crucibles and its covers were returned in the desiccators. The weighing procedure was repeated at least twice until a constant reading is obtained. After that, weight 5 g of the sample and put into each of the crucibles. Then, the crucibles together with its cover and sample were placed into the oven and were left for at least 7 hours. After at least 7 hours, the covered crucibles were taken out using a tong and it was placed directly in the

desiccators. Let the crucibles be cool for 15 minutes. After cool, the crucibles were weighed together with the cover and sample. Lastly, step 10 was repeated until the weight is constant. Determination of Ash Content First, the crucibles were placed for 1 hour in the oven at 105C. Second, the crucibles were cooled in the desiccators. Third, the crucibles were weighed rapidly and accurately. Forth, the sample was weighed (3 5g) in the crucibles. Fifth, the crucible was inserted together with the sample into the muffle furnace at 550C. Sixth, the sample was burned at least 7 hours or until no black particle present. Seventh, the crucibles and ash were cooled in the desiccators. Last but not least, the crucibles were weighed together with the ash. Determination of Crude Protein 0.15g dry sample was weighed accurately. Then, 0.8g of mixed catalyst was added. 2.5 mL of concentrated sulphuric acid was added and the tube was gently swirled to mix the content. Next, the tube was heated slowly on a heating coil under fume hood. The content was boiled until the solution becomes clear and gives blue-green colour. Boiling was continued for another 10 minutes. Flask was cooled to 40C. After that, 5mL of distilled water was added and the digested product was transferred into distillation tube. 10mL of 45% NaOH solution was added slowly to separate the two layers of solution. The distillation tube was fixed to the condenser neatly. Then, 10mL of 2% boric acid and a few drop of indicator was added in a conical flask. Conical flask was placed at the distillate platform and the tip of distillation tube was immersed into the acid solution. The content of distillation flask was mixed by swirl it gently. The ammonia solution was let being distilled into conical flask for about 120 mL. After mixing the distillation product by swirl the flask gently, unreacted boric acid was titrated with 0.05N H2SO4 until neutral.

Determination of Crude Fat First of all, the sample was weighed for 5g in the thimble and the thimble was inserted into the Soxhlet apparatus. Besides that, the round bottom flask was weighed accurately and 200 ml of petroleum ether was poured into the flask. Next, the Soxhlet apparatus was connected to the reflux and also to the round bottom flask. The reflux process was continuously proceeds for 8 hours.

After 8 hours, petroleum ether from the round bottom flask was evaporated by using rotary evaporator. After it was fully evaporated, the round bottom flask was placed in the oven with 105C for 15 minutes and was cooled down in the desiccators. Lastly, the round bottom flask was weighed on the balance with the extracted fat of batik cake in it. Determination of Crude Fibre Firstly, crucible and ashless filter paper No.541 were dried in oven for 1 hour at 105C and then cooled in the desiccators. The weight of filter paper was recorded. After that, 2g of defatted sample was weighed into 500mL conical flask. Then, 200mL boiled sulphuric acid was added and the sample was dissolved in it using spatula. Next, the conical flask was attached to the reflux condenser and the sample was boiled for 30 minutes. Meanwhile that ,the conical flask was turned every 5 minutes to make sure that the sample was always in contact with the acid. After the 30 minutes ended, the hydrolyzed mixture was filtered through filter paper Whatman No.541. Then, the residue was rinsed with boiled distilled water to remove the acid from the filtrate. Next, the residue was placed back into its conical flask, added 200mL, boiled sodium hydroxide, then attached to condenser, and boiled for another 30 minutes. After the 30 minutes ended, the hydrolyzed mixtures were later filtered through the dried and weighed filter paper. The residue was first rinsed with boiled distilled water until the filtrate was free from alkaline, followed by a small amount of alcohol, and then drained it. After that ,the residue, together with the filter paper, was placed into the crucible and dried in oven at 105C. Finally , the crucible was later be placed in muffle furnace at 550C, to be burned completely (until no black particles), then placed into the desiccators, and the constant weight was calculated. RESULTS The table below showed the results for proximate analysis of moisture, ash, crude protein, crude fat, crude fiber, and carbohydrate for our food sample (batik cake). Table 1 : Summary of proximate analysis values. Experimental value Moisture Ash Protein 6.413% 1.712% 4.94%

Fat Fibre Carbohydrate Energy

22.98% 1.995% 61.98% 1985 kJ

DISCUSSION The moisture content of a food material is defined through the following equation: % moisture = ( Mw / M sample ) x 100 Where Mw is the mass of the water and M sample is the mass of the sample. The mass of water is related to the number of water molecules. In principles, the moisture content of a food can therefore be determined accurately by measuring the number of mass of water molecules present in a known mass of sample. It is not possibly to directly measure the number of water molecules present in a sample because of the huge number of molecules involved. A number of analytical techniques commonly used to determine the moisture content of foods are based on determinations of the mass water present in a known mass of sample. In this experiment, to analyze the moisture content of batik cake, the oven drying methods are used to determine it. In oven drying methods, the mass of water is measured in a known mass of sample. The moisture content is determined by measuring the mass of a food before and after the water is removed by evaporation.. From the result, moisture content in batik cake samples is around 1.12 %. From references, the moisture content of kek batik samples from . % to . %.Kek batik is a very low moisture food. The low moisture levels in kek batik also make it a very stable food product at room temperature. As the moisture content in kek batik increases, the viscosity will also increase. This has important implications to the food manufacturer. Another important factor if there are excessive moisture can also dissolve sugar out of kek batik, redepositing it on the surface as sugar bloom. To obtain an accurate measurement of the moisture content or total solids of food using evaporation methods, it is necessary to remove all of the water molecules that we originally present in the food without changing the mass of the food. But for the low-

moisture foods such as dried fruits and vegetables, candies, chocolate, roasted coffee, oils and fats, and low-moisture foods high in sugar or protein Karl Fisher titration is more desirable. In this method only water such as bound and free water will be determined and this method is rapid only require few minutes. Principle of this method is the titrimetric determination of water is based upon the quantitative reaction of water with an anhydrous solution of sulfur dioxide and iodine in the presence of a buffer that reacts with hydrogen ions. The major disadvantage is that the water has to be accessible and easily brought into methanol solution. Many common substances, especially foods such as chocolate, release water slowly and with difficulty, and require additional efforts to reliably bring the total water content into contact with the Karl Fischer reagents. The precaution steps that we need to follow are we need to use the apparatus correctly; such as used the tong to take out the crucible from the oven, make sure the temperature of oven is 105oC and make sure the desiccators are in good condition before used it. The drying rate of test samples will be affected by the moisture conditions and number of samples in the drying device. When wet samples are placed in the drying device with nearly dry samples, completion of the drying may be restarted. The ash content is a measure of the total amount of minerals present within a food, whereas the mineral content is a measure of the amount of specific inorganic components present within a food, such as Ca, Na, K and Cl. Dry ashing is a method of ashing used in this analysis. Dry ashing procedures use a high temperature muffle furnace capable of maintaining temperatures of between 500 and 600 oC. Water and other volatile materials are vaporized and organic substances are burned in the presence of the oxygen in air to CO2, H2O and N2. Most minerals are converted to oxides, sulfates, phosphates, chlorides or silicates. Although most minerals have fairly low volatility at these high temperatures, some are volatile and may be partially lost,such as iron, lead and mercury. Three main types of analytical procedure: dry ashing, wet ashing and low temperature plasma dry ashing used to determine the ash content of foods are based on the principle that minerals have a low volatility compared to other food components and are not destroyed by heating. The choice of the methods for a particular analysis depends on the reason for carrying out the analysis, the type of food analyzed and the equipment available. For kek batik sample the suitable method of ashing is dry ashing and followed by analysis by inductively coupled plasma emission spectrometry of the specific minerals. But in this

experiment, inductively coupled plasma emission spectrometry is not been done because we are determining the ash content of the sample. For the result for ashing , average percentage is 1.712% with the deviation of 0.01. Meanwhile, coefficient variation value is 0.642 %, this value is below 5% , so this result is acceptable. The conventional dry ashing procedure is simple to carry out, is not labor intensive, requires no expensive chemicals and can be used to analyze many samples simultaneously. Nevertheless, the procedure is time-consuming and volatile minerals may be lost at the high temperatures used. Microwave instruments are capable of speeding up the process of dry ashing. Wet ashing and low temperature plasma ashing are more rapid and cause less loss of volatile minerals because samples are heated to lower temperatures. Nevertheless, the wet ashing procedure requires the use of hazardous chemicals and is labor intensive, while the plasma method requires expensive equipment and has a low sample throughput.

Determination of the ash and mineral content of foods is important for a number of reasons such as for nutritional labeling. The concentration and type of minerals present must often be stipulated on the label of a food. Ash is also important for quality. The quality of many foods depends on the concentration and type of minerals they contain, including their taste, appearance, texture and stability. Other than that, ash also important for microbiological stability. High mineral contents are sometimes used to retard the growth of certain microorganisms. Ashing is important to be determined because some minerals are essential to a healthy diet such as calcium, phosphorous, potassium and sodium whereas others can be toxic such as lead, mercury, cadmium and aluminum which is hazardous to be consume. Meanwhile during processing, ashing must done because it is often important to know the mineral content of foods during processing because this affects the physicochemical properties of foods The percentage of protein obtained was 4.94%. The standard deviation and standard error calculated were 0.042. The coefficient of variation calculated was 0.85%, which shows that the data collected was 0.85% differ from the mean calculated. The acceptable data range must be equal or less than 5.0%. Therefore, the data obtained from experiment was acceptable.

Proteins are very important molecules in our cells. They are involved in virtually all cell functions. Each protein within the body has a specific role. Some proteins are involved in structural support, while others are involved in bodily movement, or in defense against germs. Our body requires proteins for the purpose of maintenance and healthy growth. The need for consuming proteins is especially more for infants, young children, pregnant women and recovering patients. Besides, proteins also aid in the formation of antibodies that enable the body to fight infection. Proteins serve as a major energy supplier. There are distinctive kinds of proteins, each performing a unique function in the body. Protein is an essential nutrient, but some research suggests that over consuming of protein can increase the risk of developing heart disease, stroke, kidney stones and osteoporosis. The method used to determine crude protein in this experiment is micro Kjeldahl method. There are some limitations for the Kjeldahl method used. It does not give a measure of the true protein, since all nitrogen in foods is not in the form of protein. This method also has poorer precision than the biuret method. Besides, different proteins need different correction factors because they have different amino acid sequences. Furthermore, the use of concentrated sulfuric acid at high temperatures poses a considerable hazard, as does the use of some of the possible catalysts. Finally, the technique is time consuming to carry-out (http://www-unix.oit.umass.edu/~mcclemen/581Proteins.html). In this experiment the fat content of our sample kek batik was determined by using Soxhlet method. In this study we found that the crude fat content of batik cake was 22.977 %. The percentage of crude fat content in batik cake is (22.977 0.144) %. Our results showed that the crude fat content of batik cake is a little bit high. In this case, the analytical test is might not carried out correctly which lead to the standard error value is 0.102. In developed countries where food is plentiful and varied, palatability is a major determinant of food choice. Fat contributes to the palatability of foods by its texture or mouthfeel, and its flavours. All fats and oils act as carriers for fat-soluble flavour compounds. The characteristics of fats and oils also play a very important role in the manufacture and cooking of foods and in the texture and appearance of the final product. Products such as cakes or mousses need air incorporated into the mixture in order to give a well-risen texture. This is usually achieved by trapping bubbles of air in a fat/sugar mixture to form stable foam. A crumbly texture around in some pastry and biscuits are achieved by fat coating the flour particles to prevent them from absorbing water. Fat helps retain a product's moisture content

and therefore increase its shelf life and separate the layers of gluten and starch formed in the dough when making flaky or puff pastry or biscuits. The fat melts during cooking, leaving minute air pockets and the liquid present produces steam which evaporates and causes the layers to rise. Fats give a glossy appearance for example when added to hot vegetables and also add shine to sauces. Solid fats do not melt immediately but soften over a range of temperatures. Fats can be processed to rearrange the fatty acids and alter their melting point. This technology has been used to produce spreads and cheeses that will spread straight from the fridge. In deep frying the food is completely surrounded by the frying fat which acts as a very efficient heat-transfer medium. Although in healthy eating terms fat is often closely scrutinized, it is worth remembering that fat has many important functions in the body. Fat is the main energy store in the body and the most concentrated source of energy in the diet - 1g of fat provides 37kJ (9 kcal), more than double that provided by either protein or carbohydrate (4 kcal). The body's fat deposits are used to meet energy demands when dietary energy is limited, for example where people have a poor appetite or during starvation. They may also be needed when energy requirements are high such as during high levels of physical activity and for growing babies and children. As well as being an energy reserve, fat deposits cushion and protect vital organs and help insulate the body. In the diet, fat is a carrier for the fat-soluble vitamins A, D, E and K, and enables their absorption. It provides the essential fatty acids, linoleic acid (omega-6) and alpha-linolenic acid (omega-3). We all need fat. There are various advantages and disadvantages. Because of the many advantages of fat, we all require it. But there are various disadvantages as well. Lets first have a look at the advantages. Fat helps in various body functions such as nutrient absoption and nerve transmission. It is also helps in maintaining the integrity of the cell membrane. The other advantages of fat are it supplies energy and makes food taste better. There are many disadvantages as well. Fats can also create various problems. It can lead to weight gain and its also responsible for disease like cancer and also the heart disease. Therefore we can see that there are various negatives and positives of fats We can though keep on using fats without getting harmed from it. We just need to understand the difference between the good fats and the bad fats. We need to use our brain as

there are bad fats and there good fats. So what we need to do is to replace the bad fats with good fats.

As we know, the foods we eat contain fat and fat is the main source of our energy. Fat can affect the quality of foods and life. In term of physiological, fat is a source of fat-soluble vitamins, essential fatty acids, precursors of prostaglandins, and is a carrier for lipophilic drugs (Trudell, 1996). Fat is the most concentrated source of energy in the diet, which also providing proteins and carbohydrates. Fat also contributes to creaminess, appearance, palatability, texture, and lubricity of food and increases the feeling of satiety during meals (Sipahioglu, 1999). In addition, the taste, smell, mouth feel and hedonic properties of fat all contribute to the popular concept of fat taste. Fat is one of the reasons why palatability and energy density of foods are closely intertwined. Energy density of foods is largely determined by their water and fat content. Generally, the most palatable foods are those that are both energy-dense and high in fat content (Drewnowski, 1997). As in daily life, the amount of fat per serving should have a limit because excessive fat in body can lead us to an unhealthy life and cause obese and diseases such as cardiovascular diseases, high blood pressure and kidney malfunction. On the other hand, there are some forms of fat which are healthful to body as well as protect body from diseases. Monounsaturated fat is considered a good fat without any adverse effects on body. It provides protection from cancer and heart disease. Besides, another form of good fat is polyunsaturated fat. It provides all the necessary fatty acids for healthy skin and assist in the development of body cells. In addition, fat actually acts as an energy-conserve factory for the body, providing energy in times when there is no other energy source intake available. Fat deposits beneath the skin too can help insulate the body, protecting it from excessive heat or cold. Even though fat has several advantages, it is also has disadvantages. As we stated above, saturated fats should be limited and trans-fats should be avoided. Both of these types of fat will increase the risk for stroke and heart disease by increasing LDL (low density lipoprotein) cholesterol whereas trans-fats will decrease HDL (high density lipoprotein) levels in body. The weight of crude fibre was obtained from the difference between the weight of dried sample residue and the weight of ash. The average percentage of crude fibre obtained

experimentally 1.995 %. Crude fibre determinations are greatly affected by manipulations and procedures. Particles size is very vital as the finer the material is ground, the lower the determined crude fibre content. (Pomeranz and others 1971. The crude fibre data obtained can be used to measure the cellulose and lignin in a food. In addition, crude fibre data can also be used to determine the carbohydrates and energy of fond in food analysis. After the total percentage of moisture, ash, protein, fat, and fibre are calculated, the carbohydrates and energy of food can hence be calculated. However, there is a limitation from the crude fibre values. It does not determine hemicelluloses, pectin and hydrocolloids as they are digested by the alkaline and acid and consequently they are not being collected. Nevertheless, this method is a fairly simple method to carry out and is the official AOAC method for a number of different foodstuffs. Sample preparation is important as sample is used in the analysis of food. Sampling and any subsequent separations may be the greatest source error in food analysis, therefore sampling preparation must be done carefully. By sampling only a part of population, a quality estimate can be obtained accurately, quickly and with less expense and personnel time than if the total population were measured. Moreover, in the case of food products, analyzing a whole population would be practically impossible because of the destructive nature of most analytical methods. Estimated parameters using representative samples are normally more accurate than the same estimations done on the whole population. In determination of carbohydrate and energy, the percentage of carbohydrate was 61.96%. On the other hand, in order to calculate the energy, the percentage of all proximate analysis were converted to per 100g and the total of energy obtained for kek batik sample is 474.44 kcal or 1985 kJ CONCLUSION After a series of experiment had been done to determine the proximate compositions of kek batik, we are able to differentiate various method and safety on handling hazard chemical, able to understand and explain the reaction during the experiment. There are approximately 6.413%, 1.712%, 4.94%, 22.98%, 1.995%, and 61.96% for the moisture, ash, crude protein, crude fat, crude fiber, and carbohydrate contents respectively in kek batik sample. The total energy of our sample calculated is 474.4 kcal.

REFERENCES

1. Belitz, H.D. and Grosch, W. (1987). Food Chemistry. Springer-Verlag, Berlin. 2. Nielsen, S. Suzanne. (2003). Food Analysis. Third Edition. New York: Academic/ Plenum Publishers. 3. Self R. (2005). Extraction of organic analytes from foods: a manual of methods; Royal Society of Chemistry. Page 24-25 4. Pomeranz Y, Meloan CE. (1971). Food Analysis: Theory and Practice. Westport, Connecticut. 5. Dakin H.D, Dudley H.W. (1914) Some limitations of the Kjeldhal method, Journal of Biological Chemistry 6. http://global.britannica.com/EBchecked/topic/387783/moisture-content 7. Nielsen, S. Suzanne. (2003). Food Analysis, Third Edition, New York: Academic/ Plenum Publishers. 8. http://www.scribd.com/doc/24474812/Determination-of-Moisture-Content 9. http://www.dmsc.moph.go.th/webroot/drug/km/docs/Karl%20Fischer%20Titration.pd f Kluwer Kluwer

APPENDICES

Determination of moisture

1 Weight of empty dish (g) 35.135 Weight of empty dish + sample before drying (g) Weight of empty dish + sample after drying (g)

2 25.781

3 26.753

40.136

30.79

31.759

39.728

30.427

31.394

1. % Moisture Content

= = x 100

x 100

where = weight of empty dish + sample before drying

i) For sample A % Moisture content = = 8.158 % ii) For sample B % Moisture content = = 7.247% iii) For sample C % Moisture content = = 3.833% x 100 x 100 x 100

2. Mean of % of wet-weight, = =

= 6.4127%

3. Standard deviation, SD

= = 2.28

4. Coefficient of variation, CV = = = 0.356% (<5%, the value is acceptable)

5. Standard error

= =

= 1.32

Therefore, the % of moisture content is (6.4127 2.28)%

Determination of Ash Content Table 4.Summary of ash content in kek batik Weight (g) Crucible Sample Ash + crucible % ash 1 31.777 5.002 31.863 1.719 2 25.333 5.004 25.418 1.699 3 26.536 5.005 26.622 1.718

Crucible 1 % ash =

= =

= 1.719 % Crucible 2 % ash =

= =

= 1.699 % Crucible 3 % ash =

= = 1.718 %

Mean = = =

= 1.712 % Standard deviation, SD =

= = 0.011 % Therefore, the ash content in kek batik is (1.712 0.01) % Coefficient variation, CV = = = 0.642 % , acceptable Standard error = =

; CV < 5 % acceptable

= 0.006 Therefore, the % of ash is (1.712 0.011)%

Determination of Crude Protein

Titration (volume of H2SO4 used) Distillation Tube Blank A B Weight of Sample Initial (g) 0.153 0.154 (ml) 1.0 12.5 6.1 volume Final (ml) 1.2 18.6 8.2 volume Total volume (ml) 0.20 2.10 2.10

Calculation: 1. Weight of sample (g) Volume of H2SO4 to titrate Boric acid Volume of H2SO4 to titrate blank Normality of H2SO4, N Therefore, % nitrogen = % Protein = % Nitrogen x conversion factor Where, conversion factor = 5.71 Volume of H2SO4 to titrate blank, Ib = =W = Is = Ib = 0.05N

i.

For distillation tube A: % nitrogen = = 0.87% % protein = % nitrogen x conversion factor = 0.87% x 5.71 = 5.0%

ii.

For distillation tube B: % nitrogen = = 0.86%

% protein = % nitrogen x conversion factor = 0.86% x 5.71 = 4.9%

2. Mean of percentage of protein, = =

= 4.94 %

3. Standard deviation,

= = = 0.042 4. Coefficient of variation, CV= = x 100% x100%

= 0.85%

5. Standard error

= =

= 0.03

Therefore, the % of protein is (4.94 0.042)%

Determination of Crude Fat

Table. Summary of fat content of the batik cake. Weight (g) Sample, W Round bottom flask Round bottom flask + oil Oil, M % of oil in sample 1 5.001 148.785 149.929 1.144 22.875 2 5.009 98.469 99.625 1.156 23.078

Sample 1 % of oil in sample = = = 22.875 % Sample 2 % of oil in sample = = = 23.078 % Mean = = = = 22.977 % Standard deviation, SD =

= 0.144 % Therefore, the ash content in kek batik is (22.977 0.144) % Coefficient variation, CV = = = 0.627 % Standard error = =

; CV < 5 % acceptable

= 0.102 Therefore, the % of fat is (22.97 0.144)%

Determination of Crude Fibre Table 7 : weight and percentage of crude fiber of sample Sample Weight of sample, W (g) Weight of filter paper without ash, K (g) Weight of crucible + filter paper + dried residue, S (g) Weight of crucible + ash, A (g) % of crude fibre 1 2.0005 2 2.0075

0.301

0.303

50.664

46.259

50.324 1.95 %

45.915 2.04%

Mean % of crude fibre

1.995%

(S K ) A 100 W % crude fiber =

For sample 1, % of crude fibre

(50.664 0.301) 50.324 100 2.0005

= 1.95 % For sample 2, % of crude fibre =

(46.259 0.303) 45.915 100 2.0075

= 2.04% Mean % of crude fibre in food sample =

1.95 2.04 2

= 1.995% Standard deviation, =

= = 0.064 Coefficient of variation = = x 100% x 100%

= 3.108 % (less than 5%, accepted) Standard error = =

= 0.045 Therefore, the % of fibre is (1.995 0.064)%

Determination of carbohydrate: % of Carbohydrate = 100%- % moisture- %ash- % crude protein- %crude fat- %fiber = 100%- 6.413% - 1.712% - 4.94% - 22.98% - 1.995% = 61.96% Total of energy formed: Protein (g) x 4 kcal/g = 4.94g x 4 kcal/g = 19.8 kcal Carbohydrate (g) x 4 kcal/g = 61.96 g x 4 kcal/g = 247.8 kcal Fats (g) x 9 kcal/g = 22.98 g x 9 kcal/g = 206.8 kcal Total kcal = 19.8 kcal + 247.8 kcal + 206.8 kcal = 474.44 kcal 1 kcal = 4.184 kJ Total energy = 474.44 x 4.184 kJ = 1985 kJ