plus jawaban

8/10/2019 Plus Jawaban

1/33

A. Title Experiment : Phytochemical Test on Curcuma

Zanthorriza

B. Day/Date of Experiment : Tuesday/ October, 21th

2014

C.

Done : Tuesday/ October, 21th2014

D. Purpose :

1.

Selecting required tools needed based on experiment conducted

2. Selecting required materials needed based on experiment conducted

3. Identifying chemical component of plant from terpenoids, steroids,

phenolic (antraquinon, tannin, and phenol), flavonoids, and alkaloids

E. Basic Theory

Curcuma Xanthorr izaor Curcuma Zanthorr iza(Temu Lawak)

Synonym

Curcuma javanica, Curcuma zedoaria,

Javanese turmeric

.

Common name

Curcumine, ged, haridra, indian saffron,

java turmeric, jiang huang, shu gu jiang

huang, temu lawak, tumulawak.

Family

Zingiberaceae (Ginger family).

Temulawak rhizome and

Temulawak dlower
http://start%28%27tumulawak-viewt.html%27%20%29/http://start%28%27tumulawak-viewt.html%27%20%29/http://start%28%27tumulawak-viewt.html%27%20%29/http://start%28%27tumulawak-viewt.html%27%20%29/http://start%28%27tumulawak-viewt.html%27%20%29/http://start%28%27tumulawak-viewt.html%27%20%29/


2/33

Overview

Temulawak, originally from Indonesia, can grow up to 8 feet tall. The

flower is yellow. The large leaves stem from the root and the large

rhizome of the plant contains herbal qualities. Cultivation has spread to

other countries including Surinam.

Java - turmeric is a mild spice used in

several drinks to give these flavor and color

(yellow). It is also used for seasoning food

in the Indonesian cuisine; and used

extensively in India (in curry).

It has an aromatic, pungent odor and a bitter

taste.

Medicinal Applications

Interesting is that the rhizome is used medicinally; it has liver protection

properties.

Usefulness

-

The active ingredients (anti-oxidant and anti-edemic) are the

curcuminoids (curcumin, dicinnamoyl - methane, bis - demethoxy -

curcumin, demethoxy curcumin) encourage bile and prevent the

formation of gallstones.

- It also has essential oils, cinnamaldehyde and starch / carbohydrate.

The rhizomes have anti - viral and anti - inflammation properties

(Hepatitis B and C).

- Used against acne (inhibits bacterial growth); normalize digestion.It

increases breast milk production.

-

Can be used in the treatment of inflammatory bowel disease; effective

the for long-term maintenance therapy in patients with ulcerative

colitis; urinary tract infection (UTI).

-

Decreases cholesterol levels in blood and liver.
http://start%28%27view-kunirpowder02.jpg%27%29/


3/33

- A tincturecan be used against, among other applications,

high cholesteroland blood triglycerides.

-

Temu lawak can also be used as a spice for food coloring.

Hardness: USDA zone 9 B - 11.

Propagation: Rhizome and bulb.

Culture:Full sun, sandy loam soil, plant in frost free areas, can also be

grown in a pot or as a garden plant.

In the following experiment chemical component of Curcuma

Xanthorrizawould be identified using Phytochemical Test. The series of

test would discover whether Curcuma Xanthorriza contains terpenoids,

streroids, phenolic, flavonoids and alkaloids group..

Phytochemical Test

The subject of phytochemistry or plant chemistry has developed in

recent years as adistinct discipline, somewhere in between natural product

orgnic chemistry and plant biochemistry and is closely related to both. It is

concerned with the enourmous variety of organic substances that are

elaborated and accumuated by plants and deals with the chemical

structures of these substances, their biosynthesis, turnover and

metabolism, their natural distributuon and their biological function.

In all these operations, methods are needed for separation,

purification and identification of the many different constituents present in

plants. Thus, advances in our understanding of phytochemistry are directlyrelated to the successful exploitation of known techniques, and the

continuing development of new techniques to solve outstanding problems

as they appear. One of the challenges of phytochemistry is to carry out all

the above operations on vanishingly small amounts of material.

Frequently, the solution of a biological problem in, say, plant growth

regulation, in the biochemistry of plant-animal interactions, or in

understanding the origin of fossil plants depends on identifying a range of


4/33

complex chemical structures which may only be available for study in

microgram amounts.

Phytochemical progress has been aided enormously by the

development of rapid and accurate methods of screening plants for

particular chemicals and the emphasis in this book is inevitably on

chromatographic techniques. These procedures have shown that many

substances originally thought to be rather rare in occurrence are of almost

universal distribution in the plant kingdom. The importance of continuing

surveys of plants for biologically active substances needs no stressing.

Certainly, methods of preliminary detection of particular classes of

compound are discussed in some detail in the following chapters.

Although the term 'plant' is used here to refer to the plant kingdom

as a whole, there is some emphasis on higher plants and methods of

analysis Introduction 3 for micro-organisms are not dealt with in any

special detail. As a general rule, methods used with higher plants for

identifying alkaloids, amino acids, quinones and terpenoids can be applied

directly to microbial systems. In many cases, isolation is much easier,

since contaminating substances such as the tannins and the chlorophylls

are usually absent. In a few cases, it may be more difficult, due to the

resilience of the microbial cell wall and the need to use mechanical

disruption to free some of the substances present.

Methods of Extraction and Isolation

The plant material ideally, fresh plant tissues should be used for

phytochemical analysis and the material should be plunged into boilingalcohol within minutes of its collection. Sometimes, the plant under study

is not at hand and material may have to be supplied by a collector living in

another continent. In such cases, freshly picked tissue, stored dry in a

plastic bag, will usually remain in good condition for analysis during the

several days required for transport by airmail. Alternatively, plants may be

dried before extraction. If this is done, it is essential that the drying

operation is carried out under controlled conditions to avoid too many


5/33

chemical changes occurring. It should be dried as quickly as possible,

without using high temperatures, preferably in a good air draft. Once

thoroughly dried, plants can be stored before analysis for long periods of

time. Indeed, analyses for flavonoids, alkaloids, quinones and terpenoids

have been successfully carried out on herbarium plant tissue dating back

many years.

In phytochemical analysis, the botanical identity of the plants

studied must be authenticated by an acknowledged authority at some stage

in the investigation. So many mistakes over plant identity have occurred in

the past that it is essential to authenticate the material whenever reporting

new substances from plants or even known substances from new plant

sources. The identity of the material should either be beyond question (e.g.

a common species collected in the expected habitat by a field botanist) or

it should be possible for the identity to be established by a taxonomic

expert. For these reasons, it is now common practice in phytochemical

research to deposit a voucher specimen of a plant examined in a


6/33

recognized herbarium, so that future reference can be made to the plant

studied if this becomes necessary.

Extraction

The precise mode of extraction naturally depends on the texture

and water content of the plant material being extracted and on the type of

substance that is being isolated. In general, it is desirable to 'kill' the plant

tissue, i.e. prevent enzymic oxidation or hydrolysis occurring, and

plunging fresh leaf or flower tissue, suitably cut up where necessary, into

boiling ethanol is a good way of achieving this end.

Alcohol, in any case, is a good all-purpose solvent for preliminary

extraction. Subsequently, the material can be macerated in a blender and

filtered but this is only really necessary if exhaustive extraction is being

attempted. When isolating substances from green tissue, the success of the

extraction with alcohol is directly related to the extent that chlorophyll is

removed into the solvent and when the tissue debris, on repeated

extraction, is completely free of green colour, it can be assumed that all

the low molecular weight compounds have been extracted.

The classical chemical procedure for obtaining organic constituents

from dried plant tissue (heartwood, dried seeds, root, leaf) is to

continuously extract powdered material in a Soxhlet apparatus with a

range of solvents, starting in tum with ether, petroleum and chloroform (to

separate lipids and terpenoids) and then using alcohol and ethyl acetate

(for more polar compounds). This method is useful when working on the

gram scale. However, one rarely achieves complete separation ofconstituents and the same compounds may be recovered (in varying

proportions) in several fractions. The extract obtained is clarified by

filtration through celite on a water pump and is then concentrated in

vacuo. This is now usually carried out in a rotary evaporator, which will

concentrate bulky solutions down to small volumes, without bumping, at

temperatures between 30 and 40C.


7/33

In most cases, mixtures of substances will be present in the crystals

and it will then be necessary to redissolve them in a suitable solvent and

separate the constituents by chromatography. Many compounds also

remain in the mother liquor and these will also be subjected to

chromatographic fractionation. As a standard precaution against loss of

material, concentrated extracts should be stored in the refrigerator and a

trace of toluene added to prevent fungal growth. When investigating the

complete phytochemical profile of a given plant species, fractionation of a

crude extract is desirable in order to separate the main classes of

constituent from each other, prior to chromatographic analysis.

The Phenolic

The term phenolic compound

embraces a wide range of plant

substances which possess in

common an aromatic ring

bearing one or more hydroxyl

substituents. Phenolic substances

tend to be water-soluble, since

they most frequently occur

combined with sugar as

glycosides and they are usually

located in the cell vacuole.

Among the natural phenolic compounds, of which several thousand

structures are known, the flavonoids form the largest group but simplemonocyclic phenols, phenylpropanoids and phenolic quinones all exist in

considerable numbers.

Several important groups of polymeric materials in plants - the

lignins, melanins and tannins - are polyphenolic and occasional phenolic

units are encountered in proteins, alkaloids and among the terpenoids.

While the function of some classes of phenolic compound are well

established (e.g. the lignins as structural material of the cell wall; the


8/33

anthocyanins as flower pigments), the purpose of other classes is still a

matter of speculation. Flavonols, for example, appear to be important in

regulating control of growth in the pea plant (Galston, 1969) and their

adverse effects on insect feeding (Isman and Duffey, 1981) have indicated

that they may be natural resistance factors; neither of these functions,

however, has yet been firmly established.

Flavonoid Pigments

The flavonoids are all structurally derived from the parent

substance flavone, which occurs as a white mealy farina on Primula

plants, and all share a number of properties in common. Some ten classes

of flavonoid are recognized (Table 2.7) and these different classes will be

considered in more detail in subsequent sections. In this section, general

procedures of identification are discussed and methods for distinguishing

the different classes are outlined. Flavonoids are mainly water-soluble

compounds. They can be extracted with 70% ethanol and remain in the

aqueous layer, following partition of this extract with petroleum ether.

Flavonoids are phenolic and hence change in colour when treated

with base or with ammonia; thus they are easily detected on

chromatograms or in solution. Flavonoids contain conjugated aromatic

systems and thus show intense absorption bands in the UV and visible

regions of the spectrum. Finally, flavonoids are generally present in plants

bound to sugar as glycosides and anyone flavonoid aglycone may occur in

a single plant in several glycosidic combinations.

For this reason, when analysing flavonoids, it is usually better toexamine the aglycones present in hydrolysed plant extracts before

considering the complexity of glycosides that may be present in the

original extract. Flavonoids are present in all vascular plants but some

classes are more widely distributed than other; while flavones and

flavonols are universal, isoflavones and biflavonyls are found in only a

few plant families. Detailed references to the natural distribution of the


9/33

flavonoids may be found in Harborne (1967a), Harborne et al. (1975),

Harborne and Mabry (1982) and Harborne (1988, 1994).

Preliminary classification, Flavonoids are present in plants as

mixtures and it is very rare to find only a single flavonoid component in a

plant tissue. In addition, there are often mixtures of different flavonoid

classes. The coloured anthocyanins in flower petals are almost invariably

accompanied by colourles flavones or flavonols and recent research has

established that the flavones are important co-pigments, being essential for

the full expression of anthocyanin colour in floral tissues. Mixtures of


10/33

anthocyanins are also the rule, particularly in the flowers of ornamental

plants, and anyone flower tissue may contain up to ten different pigments.

Tannins

Tannins occur widely in vascular plants, their occurrence in the

angiosperms being particularly associated with woody tissues. By

definition, they have the ability to react with protein, forming stable

waterinsoluble co-polymers. Industrially, tannins are substances of plant

origin which because of their ability to cross-link with protein are capable

of transforming raw animal skins into leather. In the plant cell, the tannins

are located separately from the proteins and enzymes of the cytoplasm but

when tissue is damaged, e.g. when animals feed, the tanning reaction may

occur, making the protein less accessible to the digestive juices of the

animal.

Plant tissues high in tannin are, in fact, largely avoided by most

feeders, because of the astringent taste they impart. One of the major

functions of tannins in plants is thought to be as a barrier to herbivory.

Chemically, there are two main types of tannin, which are distributed

unevenly throughout the plant kingdom.

The condensed tannins occur almost universally in ferns and

gymnosperms and are widespread among the angiosperms, especially in

woody species. By contrast, hydrolysable tannins are limited to

dicotyledonous plants and here are only found in a relatively few families.

Both types of tannin, however, can occur together in the same plant, as

they do in oak bark and leaf. Condensed tannins or flavolans can beregarded as being formed biosynthetically by the condensation of single

catechins (or gallocatechins) to form dimers and then higher oligomers,

with carboncarbon bonds linking one flavan unit to the next by a 4-8 or 6-

8 link. Most flavolans have between two and twenty flavan units. The

name proanthocyanidin is used alternatively for condensed tannins

because on treatment with hot acid, some of the carbon--earbon linking

bonds are


11/33

plant are broken and anthocyanidin monomers are released.

Characterization thus involves the identification of the

anthocyanidin or anthocyanidins so released. Hydrolysable tannins are

mainly of two classes, the simplest being the galloylglucose depside, in

which a glucose core is surrounded by five or more galloyl ester groups.

A second type occurs where the core molecule is a dimer of gallic acid,

namely hexahydroxydiphenic acid, again with glucose attachment; on

hydrolysis, these ellagitannins yield ellagic acid. Within these two classes

it is possible to further subdivide the known compounds according to their

biogenetic origin (Haddock et al., 1982). The chemical elucidation of

tannins has been difficult to accomplish and it is only within the last

decade that some of the structural complexities have been fully

appreciated. There are, for example, significant stereochemical differences

between the proanthocyanidins of monocotyledonous and dicotyledonous

plants (Ellis et al., 1983). Detailed accounts of plant tannins include those

of Swain (1979) and Haslam (1989).

Triterpenoids and Steroids


12/33

Triterpenoids are

compounds with a

carbon skeleton

based on six

isoprene units

which are derived

biosynthetically

from the acyclic

C30 hydrocarbon,

squalene. They have

relatively complex

cyclic structures,

most being either

alcohols, aldehydes

or carboxylic acids.

They are colourless,

crystalline, often high melting, optically active substances, which are

generally difficult to characterize because of their lack of chemical

reactivity.

A widely used test is the Liebermann-Burchard reaction (acetic

anhydride- conc.H2S04), which produces a blue-green colour with most

triterpenes and sterols. Triterpenoids can be divided into at least four

groups of compounds: true triterpenes, steroids, saponins and cardiac

glycosides. The latter two groups are essentially triterpenes or steroids

which occur mainly as glycosides. There are also the steroidal alkaloids,but these are covered in this book along with the other alkaloids.

Many triterpenes are known in plants and new ones are regularly being

discovered and characterized (Connolly and Hill, 1991). So far, only a few

are known to be of widespread distribution. This is true of the pentacyclic

triterpenes a- and ~-amyrin and the derived acids, ursolic and oleanolic

acids (see Fig. 3.8). These and related compounds occur especially in the

waxy coatings of leaves and on fruits such as apple and pear and they may


13/33

serve a protective function in repelling insect and microbial attack.

Triterpenes are also found in resins and barks of trees and in latex

(Euphorbia, Hevea, etc.). Certain triterpenes are notable for their taste

properties, particularly their bitterness. Limonin, the lipid-soluble bitter

principle of Citrus fruits, is a case in point. It belongs to a series of

pentacyclic triterpenes which are bitter, known as limonoids and

quassinoids. They occur principally in the Rutaceae, Meliaceae and

Simaroubaceae and are also of chemotaxonomic interest (Waterman and

Grundon, 1983).

Another group of bitter triterpenes are the cucurbitacins, confined

mainly to the seed of various Cucurbitaceae but detected also in several

other families including the Cruciferae (Curtis and Meade, 1971). Sterols

and triterpenes are based on the cyclopentane perhydrophenanthrene ring

system. At one time, sterols were mainly considered to be animal

substances (as sex hormones, bile acids, etc.) but in recent years, an

increasing number of such compounds have been detected in plant tissues.

Indeed, three so-called 'phytosterols' are probably ubiquitous in occurrence

in higher plants: sitosterol (formerly known as B-sitosterol), stigmasterol

and campesterol. These common sterols occur both free and as simple

glucosides. A less common plant sterol is aspinasterol, an isomer of

stigmasterol found in spinach, alfalfa and senega root. Certain sterols are

confined to lower plants; one example is ergosterol, found in yeast and

many fungi.

Others occur mainly in lower plants but also appear occasionally in

higher plants, e.g. fucosterol, the main steroid of many brown algae andalso detected in the coconut. Phytosterols are structurally distinct from

animal sterols, so that recent discoveries of certain animal sterols in plant

tissues are most intriguing. One of the most remarkable is of the animal

estrogen, estrone, in date palm seed and pollen (Bennett et al., 1966).

Pomegranate seed is another source, but the amounts present are very low;

according to Dean et al. (1971), there is only 41lg estrone kg-I tissue. Less

remarkable, perhaps, is the detection of cholesterol as a trace constituent in


14/33

several higher plants, including date palm, and in a number of red algae

(Gibbons et al., 1967). Finally, the occurrence of insect moulting

hormones, the ecdysones, in plants must be mentioned, since they provide

a fascinating insight into the way plants may have evolved in order to

protect themselves from insect predation. Ecdysones were discovered in

plants in 1966 (Nakanishi et al., 1966) and have subsequently been found

in a range of plant tissues with high concentrations being present in a

number of ferns (e.g. bracken, Pteridium aquilinum) and gymnosperms

(e.g.Podocarpus nakaii).

Saponins are glycosides of both triterpenes and sterols and have

been detected in over seventy families of plants (Hostettmann and Marston

1995). They are surface-active agents with soap-like properties and can be

detected by their ability to cause foaming and to haemolyse blood cells.

The search in plants for saponins has been stimulated by the need for

readily accessible sources of sapogenins which can be converted in the

laboratory to animal sterols of therapeutic importance (e.g. cortisone,

contraceptive estrogens, etc.). Compounds that have so been used include

hecogenin from Agave and yamogenin from Dioscorea species. Saponins

are also of economic interest because of their occasional toxicity to cattle

(e.g. saponins of alfalfa) or their sweet taste (e.g. glycyrrhizin of liquorice

root). The glycosidic patterns of the saponins are often complex; many

have as many as five sugar units attached and glucuronic acid is a common

component.

The last group of triterpenoids to be considered are the cardiac

glycosides or cardenolides; here again there are many known substances,with complex mixtures occurring together in the same plant. A typical

cardiac glycoside is oleandrin, the toxin from the leaves of the oleander,

Nerium oleander, Apocynaceae. An unusual structural feature of oleandrin

(see Fig. 3.8) and many other cardenolides is the presence of special sugar

substituents, sugars indeed which are not found elsewhere in the plant

Triterpenoids and steroids 133 kingdom. Most cardiac glycosides are toxic

and many have pharmacological activity, especially as their name implies


15/33

on the heart. Rich sources are members of the Scrophulariaceae, Digitalis,

Apocynaceae, Nerium, Moraceae and Asclepiadaceae, Asclepias. Special

interest has been taken in the cardiac glycosides ofAsclepias,because they

are absorbed by Monarch butterflies feeding on these plants and are then

used by the butterflies as a protection from predation by blue Jays

(Rothschild, 1972). The butterfly is unharmed by these toxins, which on

the other hand act as a violent emetic to the bird.

Alkaloids

The identification using Culvenor-Fitzgerald. Reagent Meyer gives

peach, reagent Wagner gives brown, reagent dragendorff give white

precipitate. The alkaloids, of which some 10000 are known, comprise the

largest single class of secondary plant substances. There is no one

definition of the term 'alkaloid' which is completely satisfactory, but

alkaloids generally include 'those basic substances which contain one or

more nitrogen atoms, usually in combination as part of a cyclic system'.

Alkaloids are often toxic to man and many have dramatic physiological

activities; hence 204 Nitrogen compounds their wide use in medicine.

They are usually colourless, often optically active substances; most are

crystalline but a few (e.g. nicotine) are liquids at room temperatures. A

simple but by no means infallible test for alkaloids in fresh leaf or fruit

material is the bitter taste they often impart to the tongue.

The alkaloid quinine, for example, is one of the bitterest substances

known and is significantly bitter at a molar concentration of 1x 10-5. The

most common precursors of alkaloids are amino acids, although thebiosynthesis of most alkaloids is more complex than this statement

suggests. Chemically, alkaloids are a very heterogeneous group, ranging

from simple compounds like coniine, the major alkaloid of hemlock,

Conium maculatum, to the pentacyclic structure of strychnine, the toxin of

the Strychnos bark. Plant amines (e.g. mescaline) and purine and

pyrimidine bases (e.g. caffeine) are sometimes loosely included in the

general term alkaloid.


16/33

Many alkaloids are terpenoid in nature and some (e.g. solanine,

thesteroidal alkaloid of the potato, Solanum tuberosum) are best

considered from the biosynthetic point of view as modified terpenoids.

Others are mainly aromatic compounds (e.g. colchicine the tropolone

alkaloid of autumn crocus bulbs), containing their basic group as a side-

chain attachment. A considerable number of alkaloids are specific to one

family or to a few related plants. Hence, the names of alkaloid types are

often derived from the plant source, e.g. the Atropa or tropane alkaloids

and so on.

To illustrate their natural occurrence, some of the more common

alkaloids are mentioned (Table 5.5) in relation to their occurrence in

particular families. This table shows only those families in which over

fifty alkaloid structures have been detected (Hegnauer, 1967). These are

the angiosperm families which are particularly rich in these bases, but it

should be borne in mind that alkaloid distribution is very uneven and many

families lack them altogether.

Alkaloids are generally absent or infrequent in the gymnosperms,

ferns, mosses and lower plants. The structures of some of the more

common alkaloids are illustrated in Fig. 5.5. The functions of alkaloids in

plants are still largely obscure, although individual substances have been

reported to be involved as growth regulators or as insect repellents or

attractants. The theory that they act as a form of nitrogen storage in the

plant is not

now

generallyaccepted.


17/33

F. Tools and Materials

Tools Materials

Blender Curcumin

Ohauss balance concentrated HCl

Beaker glass 100 ml concentrated H2SO42N

Beaker glass 300 ml FeCl31%

Test tube CH3Cl

Pipette NH3

Threepod Mg plate

Drop plat methanol 60-80%

Spatula ethanol 70%

Funnel Aquadest

Reagent:

Lieberman-Burchard

Meyer

Wagner

Dragendorf

G.

Procedure

1. Methanol Curcuma Extract Preparation

Fresh Curcuma

-peeled out the skin

- sliced, dried up

-blended

Dried Curcuma Powder


18/33

2. Identification of Alkaloids using Culvener-Fitzgerald (Harborne,

1987)

5 g Dried Curcuma

Powder

-

Put into beaker glass 100 ml-Added 15 ml methanol 60-80%

heated until left 10 ml

-

Filtered using buchner funnel

FiltrateResidue

-Concentrated in

bath water

-

Used as sample

1 ml sample

-Added 1 ml CH3Cl

and 1 ml ammonia

-Put into test tube,

heated in bath water

FiltrateResidue

-

Divided into three testtube

Filtrate 1 Filtrate 2 Filtrate 3


19/33

3. Identification of Flavonoids (Harborne, 1987)

-Added 3 drops of

HCl 2N, shaken

-Let a minute until

forms layer

-

Put the upper layer

-Added 3 drops of

HCl 2N, shaken

-Let a minute until

forms layer

-

Put the upper layer

-Added 3 drops of

HCl 2N, shaken

-

Let a minute until

forms layer

-

Put the upper layer

Upper layer

-Tested with Mayer

reagent

Peachprecipitate

Upper layer

-Tested with

Wagner reagent

Brownprecipitate

Upper layer

-Tested with

Dragendorff reagent

Whiteprecipitate

1 ml sample + 3 ml ethanol 70%

-

Shaken

-

Heated, shaked

-Filtered

FiltrateResidue

-Added 0,1 g of Mg

and two drops of

concentrated HCl 2N

Red color in ethanol layer: (+)


20/33

4. Identification of Saponin (Harborne, 1987)

5.

Identification of Steroids

6. Identification of Triterpenoid

1 ml sample

-Boiled in 10 ml water

in bath water

Filtrate

-Shaken let in 15

minute, observed

Foam formed (+)

1 ml sample

-Added with 3 ml ethanol 70%, 2m l

conc, HCl and 2 m CH3COOH

Liebermann-Burchard

Color changes: Violetbluegreen

steroids (+)

1 ml sample

-Added with 2 ml CH3Cl and 3 ml

concentrated H2SO4

Color changes: sorrel in between layer

Terpenoids (+)


21/33

7. Identification of Tannin

H. Result of Experiment

1 ml sample

-Boiled in bath ater

using 20 ml water

FiltrateResidue

-

Added 2-3 drops of

FeCl31%

Greenish brown/dark blue: tannin (+)


22/33

I. Analysis

ANALYSIS AND DISCUSSION

Phytochemical test is one important step in the effort to uncover the

potential resources of herbs. Phytochemical the results of the analysis could give

clues about the existence of chemical component ( a compound ) type of class

alkaloid, flavonoid, saponin, steroids, triterpenoid, and tannin.Based on

experiment phytochemical on test extract rhizome curcuma ( curcuma

zanthorrhiza ) which aims to identify chemical component of herbs of terpenoid,

group steroids, phenolic ( anthraquinone, tannin, and phenol ) flavonoid, and an

alkaloid contained in extracts rhizome curcuma.

1.

The Preparation Of An Extract Methanol Curcuma Rhizomes

The first thing done is preparing an extract of methanol rhizomes curcuma.

Rhizomes curcuma fresh are cleaned and flayed then dried at rooms that is not

exposed to the sun, milled to get the pollen that is dry as many as five grams of

rhizomes curcuma is inserted into a beaker glass 100 mL to extracted by

immersing the powder into 15 mL of methanol 60- 80 %, and heated to speed up

the process of extracting then filtered with filters buncher.

Extract + CH3OH Yellow sample of concentrated

Filtrat produced then evaporated in water bath to produce an extract viscous. An

extract of methanol in yellow rhizomes curcuma concentrated, this is a sample for

test phytochemical on rhizomes curcuma.

2. Identification Alkaloid With Method Culvenor Fitzgerald (Harborne,

1987)

For identification an alkaloid, as many as 1 mL sample mixed with 1 mL

of chloroform and 1 mL ammoniac formed 2 layers. On the top layer of dark

sorrel colored and the lower layer of orange. Then heated above water bath then

shaken and distilled, filtrat resulting colored orange light on the top layer, and in


23/33

the lower layer of dark orange, then the top layer been divided into three part is

equally the flattened and incorporated into test tube. On tube A B C added H2SO4

2N then shaken and ignored a few minutes. On tube A then tested with reagent

meyer ( colorless ) and produced the sediment peach-colored, it can be inferred

sample positive ( + ) contain the alkaloid.

Extraction by the addition of chloroform aims to sever ties between an

acid and an alkaloid bound in tannin ionic where atoms n of an alkaloid with a

hydroxyl group bonded mutual stable genolik of the acid of tannin. On the dotted

ties with this an alkaloid shall be set free while sour tannin will be bound by

chloroform. The addition of sulphuric acid 2N it serves to bind back an alkaloid

being salt an alkaloid to be able to react with reagent heavy metal that is a specific

for an alkaloid that produces complex inorganic salts that are not soluble so as to

separate with metabolic secondary.

The addition of sulphuric acid 2N resulting in a solution of formed into

two phases because of the different levels of polarity between phases of aqueous

the polar and chloroform that relative less polar. A salt of alkaloids will dissolve

in the top layer, while a layer of chloroform was in the lining of the bottombecause it has a mass of larger kinds.While shake with strong aims to dissolve

those compounds of every layers precisely and perfect. Reagent meyer aims to

detect an alkaloid, where reagent this bind with the alkaloid through coordinate

bonds between the atoms N alkaloids and Hg reagent meyer so as to produce

complex compounds of mercury with which nonpolar settles peach- colored. The

reaction by the experiment an alkaloid with reagent meyer is:

N +K2[HgI4] NK+ +

K2[HgI4]

In tube B was tested with reagents obtained Waghner greenish brown solution and

a brown precipitate that indicate (+) test alkaloids. The reaction on the reagent test

Waghner alkaloids are:


24/33

N + KI + I2 NK+ + I3-

In tube C was tested with reagent Dragendorf light brown color and a no

precipitate is formed, that indicate (-) alkaloid test. The reaction on the reagent

test Dragendorf alkaloids are:

N + K[BiI4] NK+ + K[BiI4]

-

3.

Identification Flavonoid (Harborne, 1987)

In flavonoid test sample of 1 ml mixed with 3 mL of 70% ethanol solution

and then heated and filtered to separate the filtrate and the residue produces a red

colored solution. The filtrate plus the Mg metal, then add 2 drops of concentrated

HCl to detect the presence of flavonoid compounds will react with Mg, after the

addition of HCl, the solution turned into a brownish red. The addition of Mg metal

and HCl to detect the presence of flavonoid compounds which will react with Mg

flavonoids after the addition of concentrated hydrochloric acid with a red color

change occurs because of light absorption of flavonoids undergo changes toward

larger wavelengths due to the reduction reaction by HCl.

A compound of phenol is a compound having a ring aromatic and

containing one or two of hydroxyl groups. A compound of phenol tending readily

soluble in water because generally bonded with sugar as glycosides and usuallycontained in vacuoles of cells so that a rhizome and fronds curcuma positive there

are flavonoid. The activity of phenolic compounds in plants are many and very

diverse. Flavonoids are good reducing compound, these compounds inhibit many

oxidation reactions either in enzimatis or non enzimatis. Flavonoids act as a good

reservoir of free radical and superoxide lipid membrane so that it can protect

against the damaging reaction.

4.

Identification Saponin (Harborne, 1987)


25/33

For the identification of saponin, 1 ml sample with 10 mL of water boil in

a water bath produces orange color in the solution. After the filtrate was shaken

and allowed to stand for 15 minutes. Formed stable foam indicates positit (+) to

test for saponins saponins glikosid similar properties that have the characteristics

of skill frothy when shaken akuos solution. Saponins are components that are

ampifilik polar lipids (having hydrophilic groups and hydrophobic groups). In

liquid systems, liquid lipids spontaneously form micelles dispersed with phyllic

tails that intersect with the liquid medium. Micelles may contain thousands of

lipid molecules. Liquid lipids form a layer with a thickness of one molecule is a

single layer. In such systems, the hydrocarbon tail open so away from water and a

hydrophilic layer that extends to the water is polar, the system is called premises

foam.

Saponin is a compound of active a strong surface, give rise to foam if

shaken with water and at concentrations that are low-growing often causing

hemolysis red blood cells. Saponin in a solution very dilute, can be as a fish

poison besides saponin also reported potentially to as antimicrobial, can be used

as raw materials for the synthesis of steroid hormones. Two kinds of saponin

which was known that is a glycoside triterpenoid alcohol and glycosides structure

steroid. Aglikon called sapogenin, obtained by the hydrolysis in an acid or using

an enzyme ( robinson, 1995 ).Rhizomes curcuma containing saponin rhizomes

that is only because there are active compounds called curcumin.

5. Identification Steroids (Harborne, 1987)

On Steroids test. Samples plus 3 mL of 70% ethanol and then added 2 mL

of concentrated H2SO4and 2 mL of acetic acid and then shaken and let stand 15

minutes. The function of acetic acid is to form an acetyl derivative of the steroid.

While the function is to reduce acetyl H2SO4formed a positive test if formed blue

solution, but in this experiment formed a yellow color that indicates a negative

steroid.

6.

Identification Triterpenoid (Harborne, 1987)


26/33

Samples added 2 mL of chloroform and 3 mL H2SO4 concentrared. The

function of chloroform is to dissolve triterpenoid that it is readily soluble in

organic solvents. Function H2SO4is to reduce tripenoid that which produces color

sorrel. This exhibiting positive ( + ) the existence of triterpenoid. Triterpenoid is a

component of plants which have the odor and can be isolated from vegetable

matter by the distillation of as a volatile oil. Triterpenoid consists of a frame with

3 cyclic 6 who joined with the cyclic 5 or 4 cyclic 6 that has a cluster of on cyclic

certain ( Lenny, 2006 ).

A compound triterpenoid this test to see whether extract compound

containing triterpenoid. A compound triterpenoid will experience dehydration by

the addition of a strong acid and forms salts gave some reaction of color. A

solvent chloroform can dissolve this compound because soluble in either in

chloroform and not containing in the molecule of water. The addition of acetic

acid anhydrous will be turned into acetic acid before the reaction takes place and

are not formed a derivative of acetyl. Triterpenoid respond establishment of a

brownish color when this compound was detected of concentrated sulphuric acid

through its walls meanwhile, will produce bluish green color ( Robinson, 1995 ).

7. Identification Taninns (Edeoga et all., 2005)

Samples of 1 mL boiled with 20 mL of water and then filtered. The

resulting filtrate was added a few drops (2-3 drops) of 1% FeCl3formed brownish

black color in the solution. This indicates a negative (-) test tannins.

Test phytochemical use FeCl3to know whether an extract of brutes having

a cluster of fenol. Tanin is a compound polifenol. Bluish green color formed at

extract after added FeCl3because tannin form compound complek with FeCl3. As

a compound of phenol other tannin produces a bluish green with iron ( iii )

chloride ( Robinson, 1995 ).


27/33

The formation of color the occurrence of this because the establishment of

complex compounds between the metals Fe and tannin. Complex compounds

formed because of a covalent bond coordination between ion or atom of metal

with the atoms nonmetals ( Effendy, 2007 ). Tendency Fe in the formation of

complex compounds can fasten 6 pairs of electrons free. Ion Fe3+ in the

formation of complex compounds form will hybridization d2sp3, so will unfilled

by 6 pairs of free electrons atoms O ( Effendy, 2007 ).

The tannin stability can be achieved if repulsion between ligan on 3 tannin

minimum.This happens if 3 tannin the position taken away. The result of a

rhizome curcuma negative because does not occur the formation of the compound

colors complex between the metals Fe and tannin. Complex compounds formed

due to a covalent bond coordination between an ion or atom of metal with an atom

the nonmetals.

FeCl3 Fe3++3Cl-

HO

OH

OH

HO

OH

+Fe3+ OH

OH

HO

OH

+Fe(OH)3

black

J. Conclusion


28/33

Reference

Anonymous. 2014. Curcuma XanthorrizaTemulawak.

http://www.tropilab.com/tumulawak.html (acessed on 25 October 2014 at 9.21am)

Harborne, J.B. 1998.Phytochemical Methods A Guide to Modern Techniques of

Plant Analysis Third Edition. London: Chapman & Hall (ebook)

Philipson, J. David. 2000.Phytochemistry and medicinal plants. Phytochemistry

56 (2001) 237243 (online http://www.elsevier.comaccessed 25 October 2014 at

10 am)

Robinson, T. 1963.The Organic Constituents of Higher Plants: Their Chemistry

and Interrelationships. New York:Minneapolis, Burgess Pub. Co (ebook)

Tim Dosen Kimia Organik. 2014.Penuntun Praktikum Kimia Organik 2.

Surabaya : UNESAPRESS.
http://www.elsevier.com/http://www.elsevier.com/http://www.elsevier.com/http://www.elsevier.com/


29/33

Question and Answer

1.

Write each reaction in phytochemical test!

2.

Write basic structure of each steroids, , triterpenoids, tannin, saponin,flavonoids, and alkaloids group.

3.

Mention flavonoids compound that contained in curcuma rhizome based

on literature.

4. Mention function and benefits of curcuma rhizome for human life.

Answer

1.

Reaction

Phytochemical

test

Reagent Reaction

Identification of

alkaloids

Mayer

N +K2[HgI4]

NK+ +K2[HgI4]

Wagner

N + KI + I2

NK+ + I3-

Dragendorff

N +K[BiI4]

NK+ + K[BiI4]

-


30/33

Identification of

flavonoids

Mg+conc.H

Cl+ethanol

Identification of

saponin

Identification of

steroids

Liebermann

-Burchard

Identification of

triterpenoids

Liebermann

-Burchard

Identification of

tannin

FeCl31% FeCl3 Fe3++3Cl-

HO

OH

OH

HO

OH

+Fe3+

OH

OH

HO

OH

+Fe(OH)3

black

2. Basic structure

Alkaloids Contain N in heterocyclic ring


31/33

Flavonoids

(Robinson, 1963)

Saponin

Steroids

(Robinson, 1963)

Triterpenoids


32/33


33/33

- Used against acne (inhibits bacterial growth); normalize digestion.It

increases breast milk production.

-

Can be used in the treatment of inflammatory bowel disease; effective

the for long-term maintenance therapy in patients with ulcerative

colitis; urinary tract infection (UTI).

-

Decreases cholesterol levels in blood and liver.

- A tincturecan be used against, among other applications,

high cholesteroland blood triglycerides.

-

Temu lawak can also be used as a spice for food coloring.

plus jawaban

Documents