A.

TITLE OF EXPERIMENT
Phytochemical Testing on Red Ginger
B. DATE OF EXPERIMENT
Tuesday, 17th April 2018 at 10:20 am – 04:20 pm
C. PURPOSE OF EXPERIMENT
- Choosing the equipment that needed appropiate with experiment will be done.
- Choosing the materials that needed appropiate with experiment will be done.
- Identify the component of plants from terpenoid, steroid, fenolik (antrakuinon, tannin, and
phenol), flavonoid and alkaloid group that contain in the extract of Red Ginger Rhizome.
D. BASIC THEORIES
 Phytochemical test
Medicinal plants have been the mainstay of traditional herbal medicine amongst rural
dwellers worldwide since antiquity to date. The therapeutic use of plants certainly goes
back to the Sumerian and the Akkadian civilizations in about the third millenium BC.
Hippocrates (ca. 460–377 BC), one of the ancient authors who described medicinal
natural products of plant and animal origins, listed approximately 400 different plant
species for medicinal purposes. Natural products have been an integral part of the ancient
traditional medicine systems, e.g. Chinese, Ayurvedic and Egyptian (Sarker & Nahar,
2007). Over the years they have assumed a very central stage in modern civilization as
natural source of chemotherapy as well as amongst scientist in search for alternative
sources of drugs. About 3.4 billion people in the developing world depend on plant-based
traditional medicines. This represents about 88 per cent of the world‟s inhabitants, who
rely mainly on traditional medicine for their primary health care. According to the World
Health Organization, a medicinal plant is any plant which, in one or more of its organs,
contains substances that can be used for therapeutic purposes, or which are precursors for
chemo-pharmaceutical semi synthesis. Such a plant will have its parts including leaves,
roots, rhizomes, stems, barks, flowers, fruits, grains or seeds, employed in the control or
treatment of a disease condition and therefore contains chemical components that are
medically active. These non-nutrient plant chemical compounds or bioactive components
are often referred to as phytochemicals („phyto-„ from Greek - phyto meaning „plant‟) or
phytoconstituents and are responsible for protecting the plant against microbial infections
or infestations by pests. The study of natural products on the other hand is called

1|Phytochemical Testing on Red Ginger

 phytochemistry. Phytochemicals have been isolated and characterized from fruits such as
grapes and apples, vegetables such as broccoli and onion, spices such as turmeric,
beverages such as green tea and red wine, as well as many other sources (Doughari &
Obidah, 2008; Doughari et al., 2009).
The science of application of these indigenous or local medicinal remedies including
plants for treatment of diseases is currently called ethno pharmacology but the practice
dates back since antiquity. Ethno pharmacology has been the mainstay of traditional
medicines the entire world and currently is being integrated into mainstream medicine.
Different catalogues including De Materia Medica, Historia Plantarum, Species Plantarum
have been variously published in attempt to provide scientific information on the
medicinal uses of plants. The types of plants and methods of application vary from
locality to locality with 80% of rural dwellers relying on them as means of treating
various diseases. For example, the use of bearberry (Arctostaphylos uva-ursi) and
cranberry juice (Vaccinium macrocarpon) to treat urinary tract infections is reported in
different manuals of phytotherapy, while species such as lemon balm (Melissa
officinalis), garlic (Allium sativum) and tee tree (Melaleuca alternifolia) are described as
broad-spectrum antimicrobial agents. A single plant may be used for the treatment of
various disease conditions depending on the community. Several ailments including fever,
asthma, constipation, esophageal cancer and hypertension have been treated with
traditional medicinal plants (Cousins & Huffman, 2002; Saganuwan, 2010). The plants
are applied in different forms such as poultices, concoctions of different plant mixtures,
infusions as teas or tinctures or as component mixtures in porridges and soups
administered in different ways including oral, nasal (smoking, snoffing or steaming),
topical (lotions, oils or creams), bathing or rectal (enemas). Different plant parts and
components (roots, leaves, stem barks, flowers or their combinations, essential oils) have
been employed in the treatment of infectious pathologies in the respiratory system,
urinary tract, gastrointestinal and biliary systems, as well as on the skin (Rojas et al.,
2001; R´ıos & Recio, 2005; Adekunle & Adekunle, 2009).
Medicinal plants are increasingly gaining acceptance even among the literates in urban
settlements, probably due to the increasing inefficacy of many modern drugs used for the
control of many infections such as typhoid fever, gonorrhoea, and tuberculosis as well as
increase in resistance by several bacteria to various antibiotics and the increasing cost of

2|Phytochemical Testing on Red Ginger

 prescription drugs, for the maintenance of personal health (Levy, 1998; Van den Bogaard
et al., 2000; Smolinski et al., 2003). Unfortunately, rapid explosion in human population
has made it almost impossible for modern health facilities to meet health demands all over
the world, thus putting more demands on the use of natural herbal health remedies.
Current problems associated with the use of antibiotics, increased prevalence of multiple-
drug resistant (MDR) strains of a number of pathogenic bacteria such as methicillin
resistant Staphylococcus aureus, Helicobacter pylori, and MDR Klebsiela pneumonia has
revived the interest in plants with antimicrobial properties (Voravuthikunchai & Kitpipit,
2003). In addition, the increase in cases of opportunistic infections and the advent of
Acquired Immune Deficiency Syndrome (AIDS) patients and individuals on
immunosuppressive chemotherapy, toxicity of many antifungal and antiviral drugs has
imposed pressure on the scientific community and pharmaceutical companies to search
alternative and novel drug sources.
Extraction methods for studying phytochemicals Extraction from the plant is an
empirical exercise since different solvents are utilized at varying conditions such as time
and temperature of extraction. As bioactive components extracted from the plants further
their separation from co extractives compounds is essential. Further fractionation of
extracted compounds done on the basis of their acidity, polarity or molecular size. The
extraction methods mostly used has been discussed below:
Cold extraction method The different plants parts dried in an artificial environment at
low temperature (50-60 °C) and dried powder then further used for extraction purpose
using various solvents. Weigh the dried powder and added into conical flask with
respective solvents and allow keeping at room temperature for thirty minute shaking after
each twenty four hours for seven days. Finally filter the extract using whatman filter paper
under vacuum and dry it at room temperature in watch glass dish. Note down the weight
of each dish prior to drying of the extracts and after drying too. Calculate the weight of
the extract from the difference.
 Red ginger
Red Ginger (Z. officinale var. Rubra) is a variance of the Zingiber Officinale species
cultivated in Indonesia and Malaysia. Besides having gingerols and shogaols, it is loaded
with anthocyanin and tannin in its root bark. Red Ginger is reddish-violet in colour and
known as “Jahe Merah” by the natives. Traditionally, it is used in spices, syrup and as

3|Phytochemical Testing on Red Ginger

 remedy for rheumatism, osteoporosis, asthma & cough. Recent reports on the anti-
inflammatory effect 7-9) of anthocyanin of Z. Officinale received much attention. Oryza
Oil & Fat Chemical Co., Ltd. prompted researches into evaluating the anti-inflammatory
effect of Red Ginger with various experimental models and results revealed that aqueous
ethanol extract of Red Ginger is beneficial against acute and chronic inflammations. In
addition, human study revealed that Red Ginger Extract lowered serum hyaluronic acid,
hence prevents destruction of joint cellular matrix component. Red Ginger Extract is an
all natural analgesic & anti-inflammatory agent suitable for the application of anti-
arthritic preparations.
Physiological Compounds of RED GINGER EXTRACT
Common to the Z. Officinale family, Red Ginger Extract is rich in gingerols & shogaols
as illustrated in Figure 1. Meanwhile, [6]-gingerol and 3R,5S-[6]-gingerdiol are
characteristic compounds of Red ginger Extract due to its high concentration as compared
to other Z. Officinale species.

Figure 1

The results of proximate analysis of Zingiber officinale (ginger) is presented in Table 1. These
include those of moisture, ash, fibre, fat, protein and available carbohydrate. The rhizome of
Zingiber officinale had 28.20±0.42%, w/w of moisture, 4.20±0.01%DM of ash, the percentage
crude fibre was 10.60±0.12%, the crude protein content was 9.05±0.01%, the fat levels of
Zingiber officinale was 4.00±0.01%. The carbohydrate content showed 80.67±0.66% which is
very high on the basis of the present study.

4|Phytochemical Testing on Red Ginger

Table 2 show results of Phytochemical screening of Zingiber officinale rhizome. The results
obtained revealed the presence of alkaloids, saponins, flavonoids, polyphenols and reducing
sugars in the aqueous extracts while cardiac glycosides, saponins, flavonoids, polyphenols and
reducing sugars were present in the petroleum ether extracts (PEE).

Table 3 shows the result of mineral elements composition of Zingiber officinale rhizome. The
results obtained ranged from 0.2±0.01 mg/100 gDM for Hg to 47.60±0.17 mg/100 gDM for
calcium.

 ALKALOID IDENTIFICATION

Alkaloids, which mean alkali-like substances, are basic nitrogenous compounds of plant or
animal origin and generally possessing a marked physiological action on man or animal. The
nitrogen is usually contained in a heterocyclic ring system. According to Hegnauer‟s
classification, three main types of alkaloids are distinguished:

5|Phytochemical Testing on Red Ginger

Type of alkaloid Precursor Type of nitrogen

True alkaloid Amino acid Heterocyclic

Protoalkaloid Amino acid Non-heterocyclic

Pseudoalkaloid Non-amino acid Heterocyclic

Isolation of alkaloid
 Alkaloids occur in free or as salts of organic or inorganic acids.
 They occur together with complex mixture of water-soluble (e.g. gums and tannins) and
water insoluble compounds (e.g. resins and lipids).
 These non-alkaloidal substances interfere during isolation and purification of alkaloids.

General procedure for isolation of alkaloids from plant material:

 Alcohol 70% is used for extraction rather than 90% to dissolve both alkaloids bases and
their salts with least amounts of impurities.
 Concentrate Ammonia is the alkali of choice for basification because it is easily volatile.
 Emulsion formed during extraction can be broken by one of the following methods:
 Addition of more of either the aqueous or the organic solvent.
 Gentle stirring with glass rod.
 Heating on water bath with slight rotation.
 Addition of few drops of alc.

Detection of Alkaloids by chemical tests

Generally, alkaloids can be detected by two groups of reagents
a. Alkaloidal precipitants.
b. Alkaloidal color reagents.
Neither of these reagents is sufficiently enough for the identification of alkaloids. Both of them
together with the confirmatory (specific tests) are needed for complete identification of the
alkaloid.
a. General Alkaloidal precipitants

6|Phytochemical Testing on Red Ginger

 These tests are applied on a clean watch glass. The reagents give amorphous or crystalline
precipitates. 1 % solution of the alkaloidal salt is prepared, rendered slightly acidic with dil.
HCI.
The following are the alkaloidal precipitants used, arranged in order of decreasing
sensitivity.
1. Wagner's reagent (Iodine-potassium Iodide solution solution)
A solution of 1.3 gram iodine and 2 gram of potassium iodide in 100 ml of water. It
gives reddish brown precipitate with most of the alkaloids even the purine bases.
2. Dragendorff's reagent (potassium bismuth Iodide).
Bismuth nitrate, nitric acid, pot. Iodide and water. It gives an orange precipitate.
3. Mayer's reagent (potassium mercuric iodide).

 FLAVONOID AND PHENOLIC

Flavonoids and Phenolics Content (HPLC Analysis)
High performance liquid chromatography (HPLC) analysis of flavonoids is
present in Table 1, the concentration of the majority flavonoids (quercetin, rutin, catechin,
epicatechin and naringenin) was increased in plants when grown under 310 μmol m −2s−1.
Accumulation of the studied flavonoid components from the sink (leaves) to the source
(rhizomes) increased under low light intensity. The analysis of flavonoid components
using HPLC in leaves of ginger showed that quercetin possessed the highest
concentration, followed by cathechin. The analysis of quercetin concentration under two
light intensities indicated a higher concentration of quercetin in ginger leaves grown
under 310 μmol m−2s−1 compared with plants grown under 790 μmol m−2s−1. In addition,
quercetin decreased the photosynthesis rate through inhibition of the ATPase activity and
electron transport rate in photosynthesis photosystems
Rutin in Malaysian ginger ranged from 0.173 and 0.451 mg/g dry weight. The
highest concentration of rutin was recorded in rhizome of Halia Bentong exposed to low
light; however, when light intensity increased, accumulation of rutin in rhizome decreased
by about 31%. Rutin concentration in Halia Bentong leaves was low, especially when
exposed to high light intensity. A moderate concentration of rutin was observed in Halia
Bara rhizome under a high light level (0.324 mg/g dry weight). Rutin accumulation in
Halia Bara, although still favored in the rhizomes more than the leaves, declined in

7|Phytochemical Testing on Red Ginger

 concentration in both plant parts with increasing light intensity from 310 to 790 μmol
m−2s−1.
Catechin and epicatechin are polyphenolic antioxidant plant secondary
metabolites. The term catechin is also commonly used to refer to the related family of
flavonoids and the subgroup flavanols. Catechin concentration in Halia Bara under low
light intensity was higher in the leaves than rhizomes, which was also comparable to that
contained in the rhizomes of Halia Bentong under low light. Meanwhile, epicatechin
concentration was highest in leaves of both varieties under low light intensity (0.117–
0.118 mg/g dry weight) although the amount was not clearly different from those obtained
in the rhizome of Halia Bara under low or high light conditions. Generally, the
concentration of epicatechin in Halia Bentong was lower than that found in Halia Bara,
especially in rhizomes under high light intensity (0.078 mg/g dry weight).

Kaempferol is a rare flavonoid in plants. However, in the leaves and rhizomes of

Halia Bara and Halia Bentong, it was detected in small concentrations (between 0.042 and
0.068 mg/g dry weight).

Naringenin is a flavonoid that is considered to have a bioactive effect on human

health as an antioxidant, free radical scavenger, anti-inflammatory, carbohydrate
metabolism promoter, and immune system modulator. It is the predominant flavanone in
grapefruit and was found to have an inhibitory effect on carcinogens. Although a lack of
information has been gathered about naringenin in ginger, from the present study, its
concentration was low, ranging from 0.02 to 0.094 mg/g dry weight. Naringenin
concentration in ginger was clearly affected by the differences in varieties, light intensity,
and plant parts. Generally, Halia Bentong had a higher concentration than Halia Bara,
with more accumulation found in the leaves than in the rhizomes, especially under low
light condition. Irradiance increases leaf area-based phenolics content, which is mainly
accumulated in the epidermis Shui et al. found that ecological factors influenced
flavonoid concentration primarily during the young stage of Ginkgo biloba development;
and amongst the ecological factors studied, light and temperature had the greatest effects
on flavonoid synthesis in Ginkgo biloba.

Salicylic acid, belonging to plant phenolics group, is found in some plant species,
and its highest levels are observed in the inflorescence of thermogenic plants and in spice

8|Phytochemical Testing on Red Ginger

 herbs. Salicylic acid was not detected in gingers grown under a light intensity of 310
μmol m−2s−1. A high content of salicylic acid (0.673 mg/g dry weight) was detected from
Halia Bara leaf extract grown under 710 μmol m−2s−1 light intensity. The results of
previous studies showed that salicylic acid is capable of enhancing plant growth and
yield. Jeyakumar et al reported that salicylic acid was able to enhance the dry matter
production in blackgram. Induction of photosynthesis rate and stomatal conductance by
salicylic acid was provided in previous studies.

Nevertheless, although the leaves‟ flavonoid content is highly sensitive to biotic

and abiotic control of PAL expression the results of Waterman et al. and Mole et
al. showed parallel variations of phenolics and flavonoids under different irradiance
levels. Contrary to our results, higher phenolics content was reported in the rhizomes of Z.
officinale rather than its leaves. In addition, the results of Chan et al, reporting a high
level of flavonoid components in ginger leaves compared to the rhizomes of Z. officinale,
supported our finding. The synthesis of isoflavones and some other flavonoids is induced
when plants are infected or injured, or under low light and low nutrient condition . The
increase in soluble phenolics such as intermediates in lignin biosynthesis can reflect the
typical anatomical change induced by stressors: An increase in cell wall endurance and
the creation of physical barriers prevent walls against harmful actions However, some
plant products such as anthocyanin, cumarin and lignin are biosynthesized while phenolic
compounds are being transformed into flavonoids ,can be observed that Halia Bentong
had a higher average increase in flavonoid components (26.1%) compared to Halia
Bentong (19.5%) when the light intensity was decreased. The structure pf flavonoid is:

9|Phytochemical Testing on Red Ginger

  TERPENOID
Terpenoids (isoprenoids) encompass more than 40 000 structures and form the largest
class of all known plant metabolites. Some terpenoids have well-characterized
physiological functions that are common to most plant species. In addition, many of the
structurally diverse plant terpenoids may function in taxonomically more discrete,
specialized interactions with other organisms. Historically, specialized terpenoids,
together with alkaloids and many of the phenolics, have been referred to as secondary
metabolites. More recently, these compounds have become widely recognized,
conceptually and/or empirically, for their essential ecological functions in plant biology.
Owing to their diverse biological activities and their diverse physical and chemical
properties, terpenoid plant chemicals have been exploited by humans as traditional
biomaterials in the form of complex mixtures or in the form of more or less pure
compounds since ancient times. Plant terpenoids are widely used as industrially relevant
chemicals, including many pharmaceuticals, flavours, fragrances, pesticides and
disinfectants, and as large-volume feedstocks for chemical industries. Recently, there has
been a renaissance of awareness of plant terpenoids as a valuable biological resource for
societies that will have to become less reliant on petrochemicals. Harnessing the powers
of plant and microbial systems for production of economically valuable plant terpenoids
requires interdisciplinary and often expensive research into their chemistry, biosynthesis
and genomics, as well as metabolic and biochemical engineering. This paper provides an
overview of the formation of hemi-, mono-, sesqui- and diterpenoids in plants, and
highlights some well-established examples for these classes of terpenoids in the context of
biomaterials and biofuels. The structure of terpenoid is

 TANNIN

Tannin, also called tannic acid, any of a group of pale-yellow to light-

brown amorphous substances in the form of powder, flakes, or a spongy mass, widely
distributed in plants and used chiefly in tanningleather, dyeing fabric, making ink, and in

10 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 various medical applications. Tannin solutions are acid and have an astringent taste.
Tannin is responsible for the astringency, colour, and some of the flavour in tea. Tannins
occur normally in the roots, wood, bark, leaves, and fruit of many plants, particularly in
the bark of oak species and in sumac and myrobalan. They also occur in galls,
pathological growths resulting from insect attacks. n addition to their principal
applications in leather manufacture and dyeing, tannins are used in the clarification of
wine and beer, as a constituent to reduce viscosity of drilling mud for oil wells, and in
boiler water to prevent scale formation. Because of its styptic and astringent properties,
tannin has been used to treat tonsillitis, pharyngitis, hemorrhoids, and skin eruptions; it
has been administered internally to check diarrhea and intestinal bleeding and as
an antidote for metallic, alkaloidal, and glycosidic poisons, with which it forms insoluble
precipitates. Soluble in water, tannins form dark blue or dark green solutions with iron
salts, a property utilized in the manufacture of ink. Tannins may be classified chemically
into two main groups, hydrolyzable and condensed. Hydrolyzable tannins (decomposable
in water, with which they react to form other substances), yield various water-soluble
products, such as gallic acid and protocatechuic acid and sugars. Gallotannin, or common
tannic acid, is the best known of the hydrolyzable tannins. It is produced by extraction
with water or organic solvents from Turkish or Chinese nutgall. Tara,
the pod from Caesalpinia spinosa, a plant indigenous to Peru, contains a gallotannin
similar to that from galls and has become an important source for refined tannin and gallic
acid. The European chestnut tree (principally Castanea sativa) and the American chestnut
oak (Quercus prinus) yield hydrolyzable tannins important in leather manufacture.
Condensed tannins, the larger group, form insoluble precipitates called tanner‟s reds, or
phlobaphenes. Among the important condensed tannins are the extracts from the wood or
bark of quebracho, mangrove, and wattle. The structure of tannin is

11 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
  SAPONNIFICATION

Saponification (alkaline hydrolysis) is an important aspect of carotenoid analysis in foods

where it is particularly effective for removing colourless contaminating lipid material and
for destroying chlorophyll if present. Several different methods have been reviewed
by Schoefs (2002, 2003, 2005). Saponification also helps to solubilise large quantities of
other food components such as proteins and carbohydrates, which would otherwise
interfere with the extraction and analysis. Carotenoid esters will also be hydrolysed
and saponification is therefore to be avoided when attempting to determine esterified
carotenes such as ethyl ester of β-apo-8′-carotenoic acid (E160f) unless the objective is to
measure the free acid. The saponification procedure is usually carried out by the addition
of ethanolic or methanolic potassium hydroxide to give an overall KOH concentration
between 5% and 10% in the extraction mixture, usually in the presence of an antioxidant
such as ascorbyl palmitate. Various temperature regimes are used for the saponification,
depending on the required speed of analysis or on the requirement for minimal carotenoid
degradation. Saponification is usually followed immediately by liquid–liquid partition
into diethyl ether or a mixture of diethyl ether and hexane (or petroleum spirit) to isolate
the unsaponifiable fraction prior to solvent removal and further clean up then washed
several times with water to remove residual potassium hydroxide.
It is also important that the formation of artefacts be minimised during
extraction, saponification, and work-up (e.g. multiple cis-isomers and oxidation products).
Careful monitoring by separation methods such as TLC or HPLC is therefore necessary to
recognise any effects and to identify and measure the products (de Quirós and Costa,
2006). Enzymatic hydrolysis can be used as an alternative to saponification and can give
improved recoveries
 TEST PYTOCHEMICAL IN RED GINGER
Phytochemical analysis of phenolics, tannins, flavonoids, terpenoids, alkaloids, steroids,
saponins and glycoside as follows.
 Test for phenols and tannins. 2 mL of 2% Ferric chloride solution was added to
the crude extracts. A blue-green to black color indicates the presence of phenols
and tannis.

12 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
  Test for flavonoid. Crude extract was mixed with 2 mL of 2% sodium hydroxide
solution. An intense yellow color was formed turned to colorless on addition of
few drops of diluted acid indicated the presence of flavonoids.
 Test for steroids. Crude extract was mixed with 2 mL of chloroform and
concentrated H2SO4 was added sidewise of test tube. A red color produced in the
lower chloroform layer indicated the presence of steroids.
 Test for terpenoids. Crude extract was dissolved in 2 mL of chloroform and
evaporated to dryness. Then 2 mL of concentrated sulfuric acid was added and
heated for about 2 min. A grayish color of mixture indicated the presence of
terpenoids.
 Test for alkaloids. Crude extract was mixed with 2 mL of 1% HCl and heated
gently. Mayer‟s and Wagner‟s reagent were then added to the mixture. Turbidity
precipitate formed shows the positive result for the presence of alkaloids.
 Test for saponins. Crude extract was diluted with 5 mL of distilled water in a test
tube. The mixture was shaken vigorously. The formation of stable foam indicated
the presence of saponins.

13 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
E. TOOLS AND SUBSTANCES
TOOLS
- Ohaus balance 1 piece
- Beaker glass 100 mL 1 piece
- Beaker glass 50 mL 1 piece
- Test tube 15 pieces
- Pipettes 15 pieces
- Tripod 1 piece
- Filtered paper 5 pieces
- Spiritus bunsen 1 piece
- Rack 1 piece
SUBSTANCES
- Red ginger 10 grams
- HCl concentrated 2 mL
- H2SO4 concentrated 10 mL
- FeCl3 1 mL
- Chloroform 5 mL
- Ammonia 1 mL
- Mg metal sufficiently
- Methanol 60-80% 40 mL
- Ethanol 70% 5 mL
- Aquadest sufficiently
- Lieberman-Burchard reagent 1 mL
- Meyer reagent 1 mL
- Dragondraff reagent 1 mL
- Wagner reagent 1 Ml

14 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
F. LANES WORK
1. Preparation Methanol Extract of Sample (Red Ginger)

Red ginger powder

 Taken about 5 grams

 Entered into beaker glass 100 mL
 Soaked into 15 mL methanol 60-80%
 Separated with separatory paper

Residue Filtrate
 Using as samples

Sample

2. Identification of Alkaloid with Culvenar-Filzgerald Method

Sample

 Taken ± 1 mL
 Added 1 mL chloroform
 Added 1 mL ammonia
 Entered into test tube
 Heated on water bath
 Shaken it
 Separated it

Residue Filtrate

 Divided becoming three parts

 Entered into different test
tube

Test Test Test

Tube 1 Tube 1 Tube 1

15 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
  Added 3 drops  Added 3 drops  Added 3 drops
H2SO4 2N H2SO4 2N H2SO4 2N
 Shaken it  Shaken it  Shaken it
 Waited a few  Waited a few  Waited a few
minutes minutes minutes
 Observed until the  Observed until the  Observed until the
solution separate solution separate solution separate
each other each other each other
 Taken the upper  Taken the upper  Taken the upper
layer layer layer
 Tested with Meyer  Tested with  Tested with
reagent Wagner reagent Dragondarf
reagent

Orange Precipitate Brown Precipitate White Precipitate

(presence alkaloid) (presence alkaloid) (presence alkaloid)

3. Flavonoid Identification

Sample

 Taken ± 1 mL
 Added 3 mL ethanol 70%
 Shaken it
 Heated
 Shaken it again
 separated

Residue Filtrate

 Added 0,1 gram Mg

 Added 2 drops concentrated HCl

Red ethanol layer

(presence flavonoid)

16 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 4. Saponin Identification

Sample

 Taken ± 1 mL
 Added 10 mL water
 Boiled in steam bath

Filtrate

 Shaken it
 Waited about 15 minutes

Formed stable foam

(presence of saponin)

5. Steroid Identification

Sample

 Taken ± 1 mL
 Added 3 mL ethanol 70%
 Added 2 mL concentrated H2SO4
 Added 2 mL anhydrous acetic acid
(Lieberman-Burchard reagent

Color changing from purple into blue or

green (presence of steroid)

6. Triterpenoid Identification

Sample

 Taken ± 1 mL
 Added 2 mL chloroform
 Added 3 mL H2SO4 concentrate

Brownish red color on the surface

(presence of triterpenoid)

17 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 7. Tannin Identification

Sample

 Taken ± 1 mL
 Added with 20 mL water
 Boiled in steam bath
 separated

Residue Filtrate
 Added 2-3 drops FeCl3 1%

Greenish brown or blackish blue

(presence of tannin)

18 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 G. OBSERVATION RESULT
Observation Result
No. Experiment Procedure Hypothesis/reactions Conclusion
Before After
1 Preparation Methanol Extract of Sample (Red Ginger) Red ginger  Red Red ginger (Zingiber officinale var. Orange
powder: ginger + amarum.) contains alkaloid, flavonoid, solution, the
brown methanol triterpenoid (Kaban, Daniel, & Saleh, result of
powder = orange 2016). filtrated as
Methanol: solution extract of
colorless with red ginger.
solution precipitate
 After
doing
filtration:
orange
solution
2 Identification of Alkaloid with Culvenar-Filzgerald Sample:  Sample + Tube 1 (reaction with Meyer reagent) Red ginger
Method orange color chlorofor is positive
Chloroform: m = contains
colorless orange alkaloid
NH3: solution Tube 2 (reaction with Wagner which
colorless  Added reagent) indicates by
H2SO4 2N: NH3 = 2 the

19 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 colorless layer precipitate
Meyer (upper is that formed.
reagent: orange
colorless solution,
Tube 3 (reaction with Dragondorf
Wagner lower is
reagent)
reagent: white
orange precipitate
solution )
Dragondorf  After (Wardhana, 2016).

reagent: heating =
orange orange
solution solution
 Filtrated =
orange
solution

Tube 1
 Filtrate +
H2SO4 =
orange
solution
 Added

20 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 Meyer
reagent =
yellow
solution

Tube 2
 Filtrate +
H2SO4 =
orange
solution
 Added
Wagner
reagent =
orange
solution
with
precipitate

Tube 3
 Filtrate +
H2SO4 =
orange

21 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 solution
 Added
Dragondo
rf reagent
= orange
solution
with
precipitate
3 Flavonoid Identification Sample:  Sample + Mg(s) + 2HCl(aq)  MgCl2(aq) + H2(g) Red ginger
orange ethanol positive
solution 70% = contains
Ethanol 70%: orange flavonoid
colorless solution which
solution  After indicate by
Mg(s) = grey heated = (Wardhana, 2016). the color
powder orange changing
HCl solution into orange
concentrate:  Filtrated = (form red
colorless yellow layer).
solution solution
 Filtrate +
Mg + HCl

22 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 concentrat
e = orange
solution
(form red
layer)
4 Saponin Identification Sample:  Sample + Red ginger
orange aquadest negative
solution = pale contains
Aquadest: yellow (Wardhana, 2016). saponin
colorless  After because
heated = after 15
pale minutes the
yellow foam is
with disappeared.
bubbles
 Shaken =
foam
formed
 After 15
minutes =
foam
disappear

23 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 ed

5 Steroid Identification Sample:  Sample + Red ginger

orange ethanol = negative
solution pale contains
Ethanol: yellow steroid
colorless  Added because the
solution H2SO4 color
H2SO4 concentrat changing
concentrate: e = into
colorless blackish blackish
(Wardhana, 2016).
Anhydrous red red.
acetic acid:  Added
colorless anhydrous
acetic acid
= blackish
red

24 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
6 Triterpenoid Identification Sample:  Sample + Red ginger
colorless chlorofor positive
solution m = contains
Chloroform: orange triterpenoid
colorless solution which
solution  Sample + indicated by
H2SO4 chlorofor color
concentrate: m + (Wardhana, 2016). changing
colorless H2SO4 into
solution concentrat blackish
e = red.
blackish
red
7 Tannin Identification Sample:  Sample + Red ginger
orange aquadest is negative
solution = yellow contains
Aquadest: solution tannin
colorless  After which
FeCl3: yellow heated = + FeCl3 (aq)  indicated by
solution yellow color not
solution changing
 Added (the color

25 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 FeCl3 = still
yellow yellow).
solution

+ HCl (aq)
(Wardhana, 2016).

26 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
H. ANALYSIS
1. Preparation Methanol Extract of Samples (Red Ginger)
The first experiment is preparation methanol extract of sample that is Red Ginger.
First, prepared red ginger that form a powder because it has purpose to destroy the cell
walls are rigid so that the target compound located in the vacuole easily retrieved and
ease in testing. In this experiment we do not need to make the powder red ginger by
yourself because it may took a long time to washing, drying, blending it and etc. It will
easy with buy the powder of red ginger directly. The powder of red ginger is brown
color. After that added 10 grams of red ginger powder into beaker glass and extracted
or macerated with 30 mL of methanol 60-80% which has colorless color. The addition
of methanol make the red ginger become orange color. Extracted or maceration is
soaking process of samples using organic solvents at room temperature. This process
is particularly advantageous in the isolation of compounds of natural ingredients
because plant samples will breakdown of the cell wall and cell membrane due to the
pressure difference between the inside and outside of the cell, so the secunder
metabolite that present in the cytoplasm will be dissolved in an organic solvent and
extraction of the compound to be perfect because it can be set the long of soaking. In
this experiment methanol is a solvent widely used in the isolation process of organic
compounds of natural ingredients because it can dissolve the whole class of secondary
metabolite.
After that filtered the sampe used filteres paper, and it will get the filtrate of red
ginger that is used as sample in this experiment. The filtrate of red ginger has orange
color that will tested phytochemicals that is to identify the presence of alkaloids,
flavonoids, triterpenoids, steroids, saponin, and phenols or tanin in extract of red
ginger. Based on literature red ginger (Zingiber officinale var. amarum.) contains
alkaloid, flavonoid, triterpenoid (Kaban, Daniel, & Saleh, 2016).
2. Identification of Alkaloid with Culvenar-Filzgerald Method
The second experiment is identification the presence of alkaloid, the principle
of the alkaloid test is the sample dissolve in ammonia chloroform, to separated the
alkaloid attached to the salt (Harbone, 1987). A sample of ± 1 ml was dissolved in a
few drops of 2N sulphuric acid. The tests used three alkaloid reagents, meyer,
wagner, dragendroff reagents. The test results are positive if the Dragendorff reagent
is formed of red precipitate. Then, the formation of yellowish white sediment with the
Meyer reagent and the formation of red precipitate with Wagner reagent (Harborne in
Priyanto, 2012).
27 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 The first sample of 1 mL extract of red ginger was poured into the reaction
tube and then added 1 mL of chloroform (colorless solution) after which 1 mL of
ammonia (colorless solution) then heated over a water bath for ± 1 minute resulted in a
red brick solution. Extraction with the addition of chloroform aims to break the bond
between ionic and tannin-bound alkaloids in which the N atom of the alkaloids is
attached to each other with the phenolic hydroxyl group of tannin acid. With the
breaking of this bond the alkaloids will be free, while the tannin acid will be bound by
chloroform. After extracting, the solution was filtered and the filtrate obtained was fed
into the same three parts and added 3 drops of 2N sulfuric acid produced a red brick
solution, and shaken vigorously after it was allowed to stand for several minutes. The
addition of 2N sulfuric acid serves to bind the alkaloids back into alkaloid salts in
order to react with heavy metal reagents that are specific to the alkaloid resulting in
inorganic salt complex which is insoluble so that it is separated from its secondary
metabolic. The addition of 2N sulfuric acid resulted in the solution being formed into
two phases because of the difference in polarity between the polar aquous phase and
the less polar chloroform. The alkaloid salt dissolves on the top layer, while the
chloroform layer is at the bottom because it has a larger density. While shaking
strongly aims to dissolve compounds in each layer properly and perfectly. Separate
layers were taken up the top layer to be tested with the Meyer, Wagner and
Dragendorf reagents.
In the test with Meyer's reagent the solution yielded a yellow solution and
there is no precipitate formed, it indicated that the negative sample contained alkanoid.
In an alkaloid test with a Mayer reagent, it is thought that nitrogen in the alkaloids
reacts with K + metal ions from the tetraiodomerkurat (II) potassium to form a
potassium-alkaloid complex that settles with a yellowish white color. The reaction to
this alkaloids test by the Meyer reagent is :
HgCl2(aq) + 2KI(aq) + HgI2(aq) + 2 KCl(aq)
HgI2(aq) + 2 KI(aq) + K2[HgI4](aq)

In the test with the Wagner reagent obtained a orange solution and there is a
brown red precipitate which indicates that positive samples contain of alkaloids. In the
manufacture of Wagner reagents, iodine reacts with I-ions of potassium iodide to

28 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 produce I3 ions - which are brown in color. In the Wagner test, the K + metal ion will
form a covalent coordinate bond with nitrogen in the alkaloid to form a precipitated
potassium-alkaloid complex. Reaction to this alkanoid test with wagner reagent:

In the test with Dragendorf reagent the solution is orange and there is red
precipitate which indicates that the positive sample contains alkoloid. In the
manufacture of Dragendorff reagents, bismuth nitrate is dissolved in HCl to avoid
hydrolysis reactions because bismuth salts are easily hydrolyzed to form bismuthic
ions (BiO +), the reaction is as follows:

Reaction of bismuth hydrolysis In order for Bi3+ ions to remain in solution,

then the solution is added acid so that the equilibrium will shift to the left.
Furthermore, the Bi3 + ions from bismuth nitrate react with potassium iodide to form a
black precipitate of Bismuth (III) iodide which then dissolves in excess potassium
iodide to form potassium tetraiodobismutat
3. Flavonoid Identification
Flavonoids are a large group of phytochemicals that are protective and widely
present in fruits and vegetables. Flavonoids are often known as bioflavonoids that act
as antioxidants. The flavonoid group is polar so it is more soluble in polar and semi
polar solvents. The polarity of the compound is due to the flavonoid being a
polyhydroxy compound (having more than one hydroxyl group) (Harborne, 1987).
The formation of red color in the ethanol layer indicates the presence of flavonoids.
The polyhydroxy from flavanone will be reduced by magnesium metal in hydrochloric
acid in an ethanol solution to form a red, yellow benzopirilium salt, or called a
flavylium salt (Harborne, 1987).
As much as 1 mL of sample (orange solution) added into test tube and labeled
with “Exp. 3”, then added with ethanol 70%, no color changes. After that test tube is
shaken and heated, no color changes after heated. Then the test tube is shaken again
and filtered with filter paper, the filtrate color is yellow. After that filtrate added with

29 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 Mg and HCl concentrate, the color changes into orange solution (red layer form).
Addition of Mg and HCl metal to detect the presence of flavonoid compound in which
flavonoid will react with Mg, after addition of concentrated hydrochloric acid there is
a change in the orange top and reddish bottom solution because flavonoid undergoes
light absorption changes toward larger wavelength due to the reduction reaction by
HCl. The red color in the ethanol coating shows the presence of flavonoids in the
sample. The equation of the reaction is as follows:
Mg(s) + 2HCl(aq)  MgCl2(aq) + H2(g)

(Wardhana, 2016).
The final result is in accordance with literature (experiment conducted by Kaban, et.
All) where red ginger (Zingiber officinale var. amarum.) contains flavonoid (Kaban,
Daniel, & Saleh, 2016).
4. Saponin Identification
The fourth experiment is identify saponin in extract of red ginger. First, entered 1
mL of sample that is extract of red ginger which has orange solution into test tube.
After that added 10 mL of aquadest which colorless into test tube and the solution
color become pale yellow. The addition of water in the extract of red ginger because
saponin is compound that contain glycosides which is the glycosides have the ability
to make foam in the water that hydrolize into glucose and other compounds. Saponin
can reduce the surface tension of water, which will form a foam on the surface of the
water after shaking. Saponins will form a foam when shaken and did not disappear
with the addition of acid. After added 10 mL of aquadest, heated the sample in the
steamed bath for 5 minutes. Then shaken the solution until a foam which has white

30 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 color formed in top of solution, let the solution for 15 minutes. After that observed the
sample, and the result is a foam that formed disappeared.
Based on the result of experiment, the foam that formed will disappeared after let
for 15 minutes (the foam not stable), it means that red ginger doesn’t contain
saponin. The result is in accordance with literature where red ginger doesn‟t contain
saponin (Kaban, Daniel, & Saleh, 2016). The reaction of the test are as follows:

(Wardhana, 2016)
5. Steroid Identification
The next experiment is steroid identification experiment, which has purpose to
identify the steroid content in the red ginger. The sample that contain steroid will turn
into blackish red which is indicated that sample containing steroid. First thing that we
must do is take 1 mL of sample that is extracted of red ginger which has orange
solution and then entered into test tube. After that added 3 ml of ethanol 70% that
colorless to the sample, and the sample color change into yellow solution. Then,
added sample with 2 ml of H2SO4 concentrated and the sample become blackish red.
After it added 2 ml of acetic acid anhydrate which colorless, the sample change into
blackish red. Addition of H2SO4 concentrated and acetic acid anhydrate as reagent to
steroid test which if the sample contain with steroid will react with reagent and show
color changes that is green or blue color. And also the addition of acetic acid
anhydride will absorb water and help oxidizing acid by sulfuric acid, because acid
oxidizing reaction will not take place if water still contained there. But in this
experiment the sample changes into blackish red not become blue or green color, it
means that the red ginger doesn‟t contain stereoid.
The final result is accordance with literature (experiment conducted by Kaban, et.
All) where red ginger (Zingiber officinale var. amarum.) doesn‟t contains stereoid
(Kaban, Daniel, & Saleh, 2016).
The reaction of this experiment is:

31 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 (Wardhana, 2016).
6. Triterpenoid Identification
Test the content of triterpenoid in red ginger is based on the formation of brownish red
color on the inter-surface. First step, as much as 1 mL of sample (orange solution)
added into test tube and labeled with “exp. 6”. Then added with 2 mL of chloroform
(colorless solution), the solution color didn‟t change. After that added with 3 mL of
H2SO4 concentrate (colorless solution) from the side of test tube slowly, the solution
color changes into blackish red and the reaction tube was hot. It indicates that the
sample (red ginger) positive contains triterpenoid. The function of chloroform is to
dissolve the soluble triterpenoids in organic solvents because chloroform has semi
polar properties. The function of H2SO4 is to reduce the triterpenoid that produces a
brownish red/red color. Here is the Liebermann-Burchard reaction mechanism on the
extract sample of ginger rhizome:

32 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 (Wardhana, 2016).

The final result is in accordance with literature (experiment conducted by Kaban, et.
All) where red ginger (Zingiber officinale var. amarum.) contains triterpenoid (Kaban,
Daniel, & Saleh, 2016).

7. Tannin Identification
In this experiment 1 ml of sample added by 20 ml of water become a yellow
solution after it heated the solution to make the reaction run quickly, then was added
with 2 drops of FeCl3 1% (yellow solution (-)) resulted still yellow solution. It
indicating in the tannin is not containing in red ginger as sample. If there are a color
changing the phenolic group in the tannin compound binds to the Fe ion of FeCl3
forms a complex compound which gives a greenish brown color. The equation of the
reaction is as follows:

+ FeCl3 →
33 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 + HCl(aq)

34 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
I. CONCLUSION
Based on the experiment we can conclude that :
- In the experiments of identification of alkaloids can be done by using the Corvenor-
Fitzgerald method with several reagents ie the Mayer reagent results there is no presence
of precipitate and formed yellow solution, Wagner reagent produce red precipitate and
orange solution, or use Dragendorff reagent produce red precipitate and solution a
orange color indicating a red ginger sample identified by tube I is negative containing an
alkaloid and on tubes II and III positively containing alkanoid.
- In the flavonoid identification experiment characterized by the yield of reddish solution
on the ethanol coating after added Mg and concentrated HCl. it shows that red ginger
contain flavonoids
- Identification of saponins is characterized by the production of stable foams. In a sample
the foam formed are not stable, so it means that the red ginger negative contain of
saponin.
- Identification of steroids is characterized by blackish red color which indicated that red
ginger is negative contains steroids.
- Identification of triterpenoids is characterized by color changing into blackish red which
indicates that the red ginger positive contains triterpenoid
- Tannin identification of red ginger shows the negative result because there is no changing
in the sample.

35 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
J. REFERENCES

Harborne, J. B. (1987). Metode Fitokimia, Penuntun Cara Modern Manganalisis Tumbuhan.

(K. Padmawinata, & I. Soediro, Trans.) Bandung: ITB.
Kaban, A. N., Daniel, & Saleh, C. (2016). UJI FITOKIMIA, TOKSISITAS DAN
AKTIVITAS ANTIOKSIDAN FRAKSI n-HEKSAN DAN ETIL ASETAT
TERHADAP EKSTRAK JAHE MERAH (Zingiber officinale var. amarum.).
Jurnal Kimia Mulawarman, 14.
Wardhana, A. P. (2016). Elusidasi Struktur Senyawa Hasil Isolasi Dari Eksrtak Kloroform
Kulit Batang Tumbuhan Gowok (syzygius polycephalum) dan Uji Aktivitas
Antioksidan. State University of Surabaya, Chemistry. Surabaya: Unpublished.
Sarker, S.D. & Nahar, L. (2007). Chemistry for Pharmacy Students General, Organic and
NaturalProduct Chemistry. England: John Wiley and Sons.
Doughari, J.H. & Obidah, J.S. (2008). Antibacterial potentials of stem bark extracts of
Leptadenia lancifoli.against some pathogenic bacteria. Pharmacologyonline
Heftmann F. 1992. Chromatography: Fundamentals and Application of Chromatographic
and Electrophoretic Techniques. 5th edn., Elsevier, Amsterdam, The Netherlands.

36 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
K. ATTACHMENTS
1. QUESTION ANSWERS
1. Write down the reaction in phytochemical test
a. Alkaloid identification

Tube 1 (reaction with Meyer reagent)

Tube 2 (reaction with Wagner reagent)

Tube 3 (reaction with Dragondorf reagent)

b. Flavonoid

37 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 c. Saponification

d. Steroid

e. Triterpenoid

38 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 f. Tannin

+ FeCl3(aq) →

+ HCl(aq)

2. Write down the structure of steroid, triterpenoid, tannin, sapponin, flavonoid, and
alkanoid ?

Flavonoid Alkaloid Steroid

39 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 Triterpenoid Saponin Tanin

3. Mention any flavonoid compounds found in the ginger rhizome based on the literarur !
Most of the flavoniods present in plants, they are bound to sugar molecules as
glycosides, and in the form of mixtures. Flavones and flavonols are the most
widespread compounds of all yellow plant pigments, although the yellow color of plant
maize is usually caused by carotenoids as a characteristic of flavonoids.
4. Mention the functions and manfat rhizome temulawak for human life !
The benefits of rhizome temulawak for human life are:
- Maintaining the Function of the Heart
- Reduce Arthritis Against Cancer
- Lowering Blood Fats Overcoming
- Digestive Problems
- Breastfeeding

40 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
5. DOCUMENTATION
A. Preparation Methanol Extract Red Ginger Rhizome

10 grams of red ginger : brown powder Entered 10 grams red ginger into beaker glass

Red ginger + methanol : brownish orange Separated the mixture with paper filtered

Taken the filtrate of mixture The filtrate of red ginger : orange solution

41 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 B. Alkanoid Identification

Tools : test tube, beaker glass, rack, Materials : methanol, cloroform, ammonia,
graduated cylinder, spiritus burner, etc H2SO4, ethanol, FeCl3

1 mL red ginger sample : orange solution Red ginger + cloroform : orange solution

+ NH3 : orange solution and white + heated : orange solution

42 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 precipitate (2 layers)

+ filtered : filtrate has orange solution Divided the filtrate into 3 test tube

+ H2SO4 : orange solution Tube 1. + meyer reagent : yellow solution

43 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
Tube 2. + wagner solution : orange solution Tube 3. + dragondraff reagent : orange
with precipitate solution with precipitate

The result of alkanoid identification

C. Flavonoid Identification

1 mL red ginger sample : orange solution + ethanol : orange solution

+ heated : orange solution + filtered : the filtrate has yellow color

44 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 Filtrate + Mg : yellow solution + HCl concentrated : orange solution (form
red layer)

D. Saponin Identification

1 mL red ginger sample : orange solution Red ginger + 10 mL aquadest : pale yellow

45 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 +heated : foam formed in the top of tube Waited about 15 minutes : foam dissapeared

E. Steroid Identification

Red ginger + 3 mL ethanol : yellowish

1 mL red ginger sample : orange solution
orange

+ concentrated H2SO4 : blackish red + anhydrous acetic acid : blackish red

46 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 F. Triterpenoid Identification

1 mL red ginger sample : orange solution Red ginger + 3 mL ethanol : orange solution

+ concentrated H2SO4 : blackish red

G. Tanin Identification

1 mL red ginger sample : orange solution + 20 mL aquadest : yellow solution

47 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r
 + heated : yellow solution + 3 drops FeCl3 : yellow solution

48 | P h y t o c h e m i c a l T e s t i n g o n R e d G i n g e r