1.0 1.

Introduction Spray Dryer

A spray dryer is a device used in spray drying. It takes a liquid stream and separates the solute or suspension as a solid and the solvent into a vapor. The solid is usually collected in a drum or cyclone. The liquid input stream is sprayed through a nozzle into a hot vapor stream and vaporised. Solids form as moisture quickly leaves the droplets. A nozzle is usually used to make the droplets as small as possible, maximising heat transfer and the rate of water vaporisation. Droplet sizes can range from 20 m to 180 m depending on the nozzle. Spray dryers can dry a product very quickly compared to other methods of drying. They also turn a solution or slurry into a dried powder in a single step, which can be advantageous for profit maximization and process simplification. Spray drying involves evaporation of moisture

from an atomised feed by mixing the spray and the drying medium. The drying medium is typically air. The drying proceeds until the desired moisture content is reached in the sprayed particles and the product is then separated from the air. The mixture being sprayed can be a solvent, emulsion, suspension or dispersion.

Figure 1: Diagram of Mini Spray Dryer.

Figure 2: Functional principle of the sample feed and dispersion.

1.2

Phytochemical Constituents
Phytochemical is a natural bioactive compound found in plants, such as vegetables, fruits,

medicinal plants, flowers, leaves and roots that work with nutrients and fibers to act as an defense system against disease or more accurately, to protect against disease. Phytochemicals are divided into two groups, which are primary and secondary constituents; according to their functions in plant metabolism. Primary constituents comprise common sugars, amino acids, proteins and chlorophyll while secondary constituents consists of alkaloids, terpenoids and phenolic compounds (Krishnaiah et al., 2007) and many more such as flavonoids, tannins and so on.

1.3

Fourier Transform Infrared Analysis (FTIR)

FTIR can be used to identify chemicals from spills, paints, polymers, coatings, drugs, and

contaminants. FTIR is perhaps the most powerful tool for identifying types of chemical bonds (functional groups). The wavelength of light absorbed is characteristic of the chemical bond as can be seen in this annotated spectrum.

2.0

Objectives

The objectives of this study are; 1. To dry the liquid extract of Melastoma malabathricum leaves. 2. To identify the phytochemical constituents of Melastoma malabathricum leaves. 3. To analysis the chemical compound of Melastoma malabathricum leaves using FTIR.

3.0 3.1

Methodology Spray Dryer

2500 ml of Melastoma malabathricum extract was sprayed dried using BUCHI Mini Spray

Dryer B-191 for 3 hour at 180C for inlet temperature and 120C for outlet temperature. The powder was produced during sprayed and collected at the end of the experiment.

3.2 3.2.1

Phytochemical Test Test for Tannins

0.5 g of the powdered sample is weighed and boiled in 20 ml distilled water in a test tube.

After that, filter the solution using filter paper. 0.1% of FeCl3 is added into the sample. After that, the observation is made. If brownish green or a blue black coloration shows, there is the presence of tannins.

3.2.2

Test for Flavonoids

2 g of sample powder was put into test tube. Few drops of 1% ammonia solution, NH3 was

added into test tube contain 2 g sample powder. A yellow coloration was observed if flavonoids compound present.

3.2.3

Test for Alkaloids

5 g of powder are added into 2 ml of HCl. 1ml of Dragendroff reagent is added into the

solution. Then, the observation is made. If an orange or red precipitate produced immediately, it indicates the presence of alkaloids.

3.2.4

Test for Steroids

1 g of powder is dissolved in 10 ml of chloroform. Next, 10 ml of sulphuric acid is added into

the solution. If the upper layer turns red and sulphuric acid layer showed yellow with green fluorescent, it indicates the presence of steroids.

3.2.5

Test for Terpenoids

5 g of powder is mixed with 2 ml of CHCl3 in a test tube. Concentrated sulphuric acid is

added carefully into the mixture to form a layer. Observation is made. An interface with reddish brown coloration is formed if there is the presence of terpenoids.

3.3 3.3.1

Production of Pellet Herbal Extract & KBr Pellet Press Preparation

The herbal extract and KBr crystal was weighted about 0.1204 g and 0.0811 g using

analytical balance. Then, mix the herbal extract with KBr crystal and grinded in the special mortar and pestle until its shiny and smooth. The purpose to grind this mixture was to reduce the particle size. The mixture need to be avoided from exposed to the air because KBr is a hygroscopic and will absorb moisture easily.

3.3.2

Pellet Preparation using KBr Mini Press

One dry bolt was placed into the dry barrel and advance five full turns. Half of the mixture

was added to the press and tapped gently to spread the sample uniformly over the lower bolt. Screwed one of the bolts halfway in the press. Then, screw in the other bolt and finger tighten. The sample was leaved under pressure with press the solid cylindrical bar until it reached 10 tons. After it reached 10 tons, loosed the bolt and removed the pellet.

3.4

Analysis of Fourier Transforms Infrared Spectroscopy (FTIR)

3.4.1

Obtain the Spectrum using FTIR

The sample was placed appropriately in the holder in the sample chamber. Make sure the

sample was squarely in-line with the source (a red laser beam should be visible through the sample). Pressed the survey softkey and make sure that the scale and the range are correct. End the experiment when the peak was already shown. Place a new sheet chart paper on the plotter. Spectrum has been plotted. Ensure the acceptable spectrum. Removed the sample and cleaned up the KBr mini press.

4.0 4.1

Result and Discussion Spray Drying

Spray drying is a technical method used to dry aqueous or organic solution. Spray drying is

involved evaporation of moisture from an atomised feed by mixing the spray and the drying medium. A liquid sample will dried and changed into powder sample. In this experiment, BUCHI Mini Spray Dryer B-191 was used. Its principle is same with co-current flow atomizer which means the sprayed product and drying air flow are in the same direction. Melastoma malabathricum was used as a sample that needs to dry by change it in powder form.

Based on Table 1, 2500 ml water extract of Melastoma malabathricum was dried and the weight of powder that produced is 17.8 g. The amount of powder form produced after sprayed is depending on the total solids content in the sample. The larger amount of total solids content in sample, the larger amount of powder produced at the end of spray drying. Meanwhile, a particle size of the final product can be influenced by the spray flowrate setting. The higher the spray flowrate will cause the smaller the size of the particles in the final product. In fact, the spray concentration influences the particles size. Therefore, the higher the concentration of the spray solution can produce the larger and the more porous the dried particles. The particles size of Melastoma malabathricum powder in this experiment is smaller which means it has less concentration of the spray solution.

Then, a powder of Melastoma malabathricum was going a phytochemical test and FTIR analysis in order to determine the components in Melastoma malabathricum leaves.

Table 1: Data of spray dryer. Item

Water extract of Melastoma sp.

Volume spray (ml)

2500

Weight or powder (g)

17.8

4.2

Phytochemical Test

Phytochemical screening was run in order to determine the constituents in Melastoma malabathricum leaves while identify either it has changes before and after spray or not. The difference for phytochemical test after spray is in powder form while before spray, a test was done in liquid form. Both samples are extracted by using water but difference equipment extraction was used.

A water extract which in liquid form was extracted by direct heating or dipping method. It was soaked in the beaker while applied a heat and stirred the solution. It is suitable because of the smaller amount of ground Melastoma malabathricum leaves. Compared to sample in powder form that produced after spray, it was extracted by using hydrodistillation. Hydrodistillation is one of the extraction methods that can apply when the larger amount of sample need to extract.

In this experiment, Melastoma malabathricum leaves are in larger amount, so it was extracted successfully by using hydrodistillation. 1.29 kg of Melastoma malabathricum leaves was extracted by mixed with 10 L of water. The process of extraction was stopped after 2 hours and by observing the colour change in mixture. The clear liquid was become yellow greenish which indicate the releasing of active ingredients in senduduk.

Table 2: Test for preliminary phytochemical screening before and after spray drying. Test for phytoconstituents
Test for flavonoids Test for tannins Test for carbohydrates Test for alkaloids Test for steroids Test for terpenoids * W - Water extract + Present

Before spray drying (W)

+ + + + + + x Not test

After spray drying (W)

+ + x + + +

Based on Table 2, a comparison before and after spray drying for phytochemical test can be made. Table 2 shows that flavonoids, tannins, alkaloids, steroids and terpenoids shown positive sign which means it is released after extraction process using water as solvent. There are no changes of these constituents when evaluation on sample was made before and after spray drying even though in difference form. Test on carbohydrates was not run due to out of stock of chemical.

Meanwhile, Table 3 shows the criteria that appeared during phytochemical test. These criteria was referred the presence of constituents in senduduk leaves. When phytochemical test was run, it used a difference of chemical that actually can react with powder of senduduk leaves extract. Then, if the constituents were detected in mixture of powder, changes of colour occurred. It was presented the type of constituents presence in Melastoma.

T bl TE T

The phyt

hemi l test for Mel t

OB E


Al aloid

y y y y y y y y

Red precipitate formed. Indicates presence of al aloid. 3 layers formed. First layer is clear, second layer is cloudy red and third layer is green fluorescent. Indicates presence of steroids 3 layers formed. First layer is reddish brown, second layer is cloudy brown and third layer is clear (colourless). Reddish brown indicates presence of terpenoids. Formation of brownish green. Indicates presence of tannin. Two layers formed. First layer is dark reddish and second layer is light brown. No flavonoids compound presence.

Steroids

Terpenoids

Tannin

y y y y y

Flavonoid

Reddish brown

Fi

3: Test for terpenoids before spray.

 l t i

powder extracts.

T ON

Fi

4: Test for terpenoids after spray.

Figure 3 and Figure

and after spray drying. Both sample was shown the presence of terpenoids when a reddish brown colour appeared after testing. Water extract sample before spray drying shows the colour is more on brown even it still has red. Meanwhile, the senduduk powder was clearly showed the cha nges of colour to reddish brown.

Fi

5: Test for steroids before spray.

Fi

7: Test for alkaloids before spray.

shows the result of terpenoids test at difference stage which is before

Three layers

Fi

6: Test for steroids after spray.

Red precipitate

Orange precipitate

Fi

8: Test for alkaloids after spray.

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Then, testing of steroids and alkaloids on senduduk powder was run. For steroids, it was shown in Figure  that it has three layers at the end of testing. Based on Figure , first layer shows a   

clear li uid, second layer is cloudy red and third layer shows the green fluorescent. These three layers showed the presence of steroids. When compared to Fi ure 5 which is testing done before spray g drying, it has more concentrated of cloudy red at second layer. Overall, it has three layers that is not differs to Figure . Figure 7 and 8 shows a difference colour that produced during testing of alkaloids. Orange precipitate was produced in Figure 7 but red precipitate only appeared in Figure 8. The production of orange or red precipitate in sample mixture actually shows the presence of alkaloids. The difference colour in alkaloids test occurred due to the difference form of sample that being tested.

Blue black coloration

Brownish green

Fi

9: Test for tannins before spray.

Fi

10: Test for tannins after spray.

Figure 9 and 10 shows the result of tannins test for water extract and powder form of senduduk extract. After tested on senduduk powder, it produced a brownish green colour in the sample. This indicates tannins were presented in the sample. It is difference when compared to result of water extract which has produced blue black coloration. But, the tannins still presence in both samples even it was differed in terms of sample form. Result for flavonoids after spray drying shows the presence of flavonoids by observed the appearing of dark reddish coloration. Dark reddish coloration was appeared in both samples by looking at Figure 11 and 12.

Dark reddish

Fi

11: Test for flavonoids before spray.

Fi

12: Test for flavonoids after spray.

4.3

Pellet

T ble 4: Spectrum of pellet. Regi li t 401 (Hydroxy compound)

P60 A50

3650-3595 2300-2000

FTIR analysis was run to determine the active compounds in the senduduk powder and compare with the result of phytochemical test. This analysis was run by prepared a pellet first. Pellet was prepared by mixed 0.120 g of potassium bromide crystal (KBr) with 0.0811 g of senduduk powder. KBr, which is a white powder, become a hard, solid, transparent pellet when subjected to high pressure. It also does not absorb infrared radiation in the same regions where organic compounds commonly absorb. The finely powdered potassium bromide will absorb more humidity from the air and therefore lead to an increased background in certain ranges. Therefore, the grind process needs to do in fast. The sample must be very finely ground to reduce scattering losses and absorption band distortions. The concentration of the sample in KBr should be in the range of 0.2% to 1%. The pellet is much thicker than a li uid film; hence a lower concentration in the sample is required. Too high a concentration usually causes difficulties obtaining clear pellets.  

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Then, the pellet was prepared by using KBr Mini Press. After pressed, a pellet was produced in thin size. Pellet was analyzed using FTIR. The thinner of pellet size cause the sample absorb beam easily. Based on the graph of FTIR analysis, two regions were detected the presence of hydroxyl compound which are 3854.440 and 3568.206.

5.0

Conclusion
Drying process of water extract of Melastoma malabathricum by using BUCHI Mini Spray

Dryer B-191 was successful done by changed a liquid form into powder form. 2500 ml water extract of senduduk was sprayed to produce 17.8 g of powder. The powder of senduduk extract was testing its phytochemical constituents in order to compare the releasing of active compound after spray with result before spray drying. Phytochemical test was shown the presence of alkaloids, flavonoids, terpenoids, steroids and tannins. All constituents that presence before spray also presence after spray even the form was changed from liquid to powder.

Then, a pellet was prepared by using KBr Mini Press. The pellet was prepared by grinding mixture of senduduk with KBr and then it was going to compress to produce a thin pellet. By analysis of pellet using FTIR, a hydroxyl group was identified. The identification of chemical compound in herbal is depending on the solvent that being used.

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6.0

References

1. Krishnaiah, D., Sarbatly, R. and Bono, A. (2007). Phytochemical antioxidants for health and medicine A move towards nature. Biotechnology and Molecular Biology Review. 1(4): 097104. Academic Journals.

2. BUCHI Labortechnik AG. (1997). Training Papers Spray Drying (Version B).

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