Analytical

this is my final assignment in  analytical chemistry. hope that it will be useful for you;D

SEPARATION OF ALKALOID IN JOHAR LEAF’S (Cassia siamea Lamk) USING EXTRACTION

I.            INTRODUCTION

Indonesia is a country with tropical climate. As a consequence, Indonesia has rich and varied flora. Local people have a long tradition of using plants for medicinal purposes and they are still utilising them. However, most of the plants have not been studied chemically and pharmacologically, and there is great potential for the isolation of novel, maturally-occuring bioactive compounds.

The leaves of Cassia siamea Lamk or commonly named by daun johar had been used as plant medicine, especially traditional drug. It is usually used by some people to overcome malaria diseases. Traditional use of Johar leaf againts malaria disease may be effective to kill the malaria parasites. It also can used to wash the injury and incentive the appetite. Based on the research Cassia siamea leaves contain sitosterol, lignin, para kumarat acid, kuinon, and alkaloid (Astuti, 2010).

One kind of separation method can use to separate alkanoid from Johar leaf’s is extraction. Basically, extraction used the difference of solubility of solute in two liquids phase which doesn’t soluble each other. Generally extraction is a attraction process of a solute in its solution in water by the other solvent which doesn’t dissolve in water. In this case, we will separate alkaloid from Johar (Cassia siamea) leaf.  

II.            DESCRIPTION OF STUDY AREA

The leaf of Cassia siamea Lamk usually used by some people to overcome malaria diseases. Traditional use of johar leaf againts malaria disease may be effective to kill the malaria parasites or to reduce fever, because people who are suffering from malaria produce fever. Malaria, caused by parasites of the genus Plasmodium, is one of the leading infectious diseases in many tropical regions (Adebayo and Krettli, 2010). It also can used to wash the injury and incentive the appetite.

Alkaloids have diverse structures and many show a range of pharmacological activities including antimicrobial activity. They are also normally readily separable from the other plant metabolites as a result of their basicity (Hadi and Bremmer, 2001). Almost alkaloid has certain pharmacological reactivity. Some alkaloid is poisoning, and the other is very useful in medical treatment. In human, alkaloid is used to reduce the fever, painful, blood pressure and tumor activity (Henry in Astuti, 2010). In plant, alkaloid is used to metabolism, self protection from insects, parasite, fungi, and bacteri. It also used to reduce or accelerate the growth (Schlitter in Astuti, 2010)

Most of alkaloid has polycyclic skeleton which contain atom C, O, N, H, and the other. This compound can be found in  solid state, amorf, and almost colourless solution. Alkaloid with complex structure and aromatic has colour, like berberine and betanine has yellow colour (Henry in Astuti, 2010).Generally alkaloid is base and soluble in organic solvent. This base properties based on the existence of lone pair in functional group of nitrogen which bond with nitrogen, and it’s aromaticity (Achmad in Astuti, 2010)

                         Picture 1: Molecular structure of alkaloid

III.            METHODE OF EXPERIMENT

Based on the literature, materials and method used is:

A.    Equipments

1.      Beaker glass

2.      Mortar and pestle

3.      Filter paper

4.      Evaporating dish

5.      Oven

B.     Materials

1.      Cassia Senea Lamk leaf

2.      Methanol

3.      Acetic acid 5%

4.      Chloroform

5.      Water(aquades)

6.      Sodium carbonate

C.     Method

1.      100 gram of Johar leaf are grinded by mortar and pestle.

2.      Soak or submerged in methanol.

3.      Filter the filtrate by filter paper

4.      Evaporate in temperature 40⁰C until dry

5.      Dried in oven in temperature 40⁰C as long 48 hours

6.      Added by acetic acid 5% until produce alkaloid salt which is soluble in water, the colour of solution turn brown.

7.      Extract the alkaloid salt by chloroform until get the colourless solution in chloroform, and solution in water phase is brown.

8.      Added sodium carbonate to attract the alkaloid salt until PH is 10 and the colour is red brownish.

9.      Extract again by chloroform until there is no alkaloid in water.

10.  Evaporate the organic solvent (chloroform) until get the brown precipitate.

IV.            DATA ANALYSIS

In the second procedure, we soak or submerged the Johar leaf powder to dissolve all the organic compound contained in this powder in organic compound, methanol. Extract of methanol got is filtered to avoid the big particle of solution. This extract is green brownish. Then we evaporate it in oven and in temperature 40⁰C in order to evaporate the methanol. The temperature is quite low because alkaloid is easy to damage in high temperature. 

The extract contains many organic compound. To separate the organic compound from alkaloid we used acetic acid 5%. Alkaloid will react with acetic acid produce alkaloid salt, and the other organic compound will react with acetic acid to produce organic salt. The colour of solution changes into brown. Alkaloid is polar, so it can soluble in water. The organic salt will soluble in organic compound, so we can separate the organic salt and alkaloid salt by the difference of solubility in water and organic compound.

The organic solvent use to separate the alkaloid salt and organic salt is chloroform. Organic salt will soluble in chloroform, and the alkaloid salt will soluble in water. This extraction is doing several times until the colour in organic phase is colourless, and the colour in water phase is brown. That means all of the organic salt is soluble in chloroform and alkaloid salt soluble in water phase.

To attract the alkaloid from its salt, we add base because adding of base can form pure alkaloid. Base we used is sodium carbonate. We add sodium carbonate until the pH is 10 because most of alkaloid can be extracted in this pH. The colour of this solution is red brownish.

This solution will be extract again by organic solvent. Organic solvent used is chloroform, because alkaloid (not alkaloid salt) will soluble in organic solvent. From the last extraction, we will get the pure alkaloid. After extraction, the organic phase is evaporated in temperature 40⁰C. The alkaloid extract is brown.

V.            CONCLUSION

In this extraction of alkaloid in Johar leaf or Cassia siamea Lamk, we use methanol, chloroform and water as the solvent. Methanol is used to extract all of the organic compound. Chloroform is used to extract the organic salt and pure alkaloid. Water is used to extract the alkaloid salt. Acetic acid is added to produce the organic salt and alkaloid salt which can separate by chloroform. Adding of Sodium carbonate is in order to reach pH 10, so all of pure alkaloid can be attracted from it salt.

VI.            REFRRENCES

Adebayo, J. O., and Krettli, A.U,. 2011. Potential antimalarials from Nigerian Plants: A review. Journal of Ethnopharmacology, vol 133, issue 2, pages 289-302.

Astuti, Yuni. 2010. Isolasi, Identifikasi, dan Karakterisasi Alkaloid dari Kulit Kayu Johar (Cassia siamea Lamk)(pdf).(Online). http://repository.ipb.ac.id/bitstream/handle/123456789/26027/Yuni%20Astuti%20(TM).pdf accesed on 17 december 2011.

Hadi, Surya and Bremmer, John B,. 2001. Initial Studies on Alkaloids from Lombok Medicinal Plants (pdf). Wollongong: Department of Chemistry, University of Wollongong, Australia.

Jumpatong, Kanlaya., Phutdawong, Weerachai., Buddhasukh, Duang. 2006. Dechlorophyllation by Electrocoagulation (pdf). Molecules 2006,11,156-162.

Riza, Fauziyah. Isolation and Identification the chemical Component from Active Mucolytic Fraction of Johar Leaf (Cassia siamea Lamk). Bandung: IPB.