Research Article

Parameters of quality control of ganda (Allium odorum L.)

leaf extract
Yoppi Iskandar1, Resmi Mustarichie2*

ABSTRACT

Aim: The aim of this research was to determine quality control parameters daun ganda (Allium odorum L.) leaves extract.
Material and Methods: Ganda (A. odorum) leaves were collected from Monaco plantations, Lembang, West Java and
determined at the Taxonomy Laboratory, Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas
Padjadjaran. Methods included These parameters include extract specific gravity, drying losses, moisture content, volatile
oil content, total ash content, acid insoluble ash content, water-soluble compounds, ethanol soluble compounds, thin-layer
chromatography, and gas chromatography-mass spectrometry (GC-MS). Results: The results of the analysis were as follows:
Extract specific gravity 1.19–2.16, drying losses 7.06–7.75%, moisture content 11.9–14%, essential oil content 0.06%, total
ash content 4–6.4%, acid insoluble ash content 0.7–1.2%, levels of water-soluble compounds 9.12–12.9%, and levels of
ethanol soluble compounds 13.6–13.75%. The results of phytochemical screening showed that the extract contained alkaloid
compounds, flavonoids, steroids, polyphenols, quinones, and monoterpenoid/sesquiterpenoid. The results of GC-MS show
that the extract contains compounds that have molecular weights 167, 207, 467, 495, and 496 and other undetectable
compounds. Conclusion: The results of this determination could be used as a preliminary description of the quality of ganda
leaf extract quality.

KEY WORDS: Allium odorum, Daun ganda, Extract parameter, Quality control

INTRODUCTION Nepal, Thailand, and the Philippines are traditionally

Allium odorum is thought to originate from China,
spread to Japan, Korea, India, Nepal, Thailand, and
the Philippines. It is an annual plant that grows in
clumps, leaves the width of 5–10  mm, solids and
slightly sprouts, leaves reach 20 cm long, dark green,
distinctive aroma, 40 cm tall flower stalks, white color,
grow upright, solid flower stalks, and square.[1,2] This
plant is known in Indonesia with various regional
names such as singando (Palembang), chives or
doubles (Sundanese), kecai, kucai, pucai (Javanese),
bucay (Madura), ganda (Minahasa), bring iosina
(Gorontalo), kocai (Bugis), kusai (Roti), and ganda
(Halmahera and Ternate).[3] Through this paper, it
will be called as ganda leaf. The usefulness of kucai
leaves is not yet widely known. However, there are
several uses that have been reported based on several
libraries as follows: In Japan, China, Korea, India,
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known to be effective in restoring fatigue.[1] The
leaves are commonly used as spices that can be
consumed in both fresh and dry forms. Clinically,
this plant is used as an anti-tumor, digestive disorder
in the intestine. In Thailand, the seeds are used in
the treatment of a toothache.[4] In another literature,
it is stated that ganda leaves have antibacterial and
antifungal activity both for Gram-positive and Gram-
negative bacteria.[5,6] It also reported as antioxidant,
anti-inflammation, antiplatelet, anti-allergic, and
cytotoxicity, reduce the risk for heart disease or
cancer.[7]
Ganda leaves contain sodium, potassium, calcium,
phosphorus, magnesium, manganese, Vitamins A,
B1, B2, C, sulfur compounds, quercetin-3-glycosides,
glucose, galactose, ferulic acid, p-kumarat acid, malic
acid, citric acid, palmitic acid, linoleic acid, and
linolenic acid.[1,8]
So far, the information about the parameters used to
determine the quality of double leaf extract has not

Department of Biology Pharmacy, Faculty of Pharmacy, Universitas Padjadjaran, Indonesia, 2Department of Pharmaceutical
1

Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, Indonesia

*Corresponding author: Resmi Mustarichie, Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of
Pharmacy, Universitas Padjadjaran, Indonesia. E-mail: *resmi.mustarichie@unpad.ac.id

Received on: 21-12-2018; Revised on: 18-01-2019; Accepted on: 14-02-2019

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 Yoppi Iskandar and Resmi Mustarichie
been widely disclosed. On this basis, the idea arose to
conduct research on the parameters used for quality
testing of ganda leaf extract. The parameters for
determining the quality of double extracts need to be
known to complete information about these plants.
MATERIALS AND METHODS
Materials
Ganda (A. odorum) leaves were collected from
Monaco plantations, Lembang, West Java and
determined at the taxonomy Laboratory, Department
of Biology, Faculty of Mathematics and Natural
Sciences, Universitas Padjadjaran. Chemical used was
Dragendorff reagent (mixture of solution Bi (NO3)3.
H2O in HNO3), Lieberman-Burchard reagent (mixture
of anhydrous acetic acid and concentrated sulfuric
acid), Mayer reagent (mixture of HgCl2 solution in
water and KI in water), magnesium (Mg) powder (CV
Agung Menara Abdi), vanillin sulfate, chloroform,
ethyl acetate, and toluene (PT Brataco). Unless stated
otherwise, all chemicals were analytical grades.
Equipment
The equipment used in this study was macerator, mortar,
stamper, drip plate, water bath, 254  nm ultraviolet
(UV) lamp and 366 nm lamps, gas chromatography-
mass spectrometry (GC-MS) (Shimadzu GC-17 A;
GC/MS. QP5050A), microscope (Nixon), digital
cameras (Nikon), and ovens (Memmert).
Methods
A collection, determination, and processing of
materials: Ganda leaves were collected and then
cleaned from the soil, washed using running water and
then dried. Then, sliced or chopped with ± 3 cm long
and dried with indirect sunlight for about 2  weeks.
After drying, the sample was ground until smooth,
then stored in a clean and closed container.[9,10]
Extraction
The extraction method used in this study was
maceration. The selection of this method was
done to prevent the occurrence of damage to the
thermolabile chemical compounds contained in
the ganda leaves leaf. The maceration was carried
out by soaking the sample in the macerator then
leaving it for 24 h at room temperature with stirring
occasionally. The solvent replacement was carried
out during 3 × 24 h.[11-13]
Examination of Quality Control Parameters from
the Extract of A. odorum
These examinations included organoleptic,
macroscopic, microscopic, phytochemical screening,
non-specific parameter, examination of specific
parameters, thin-layer chromatography (TLC), and
GC-MS referring to official literature.[9-11,14]
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Organoleptic Examination
Macroscopic and organoleptic examinations performed
on sample included physical characteristics such as
size, color, surface characteristics, texture, friability,
and the surface of the fault or the plane.[15]
Microscopic Examination
The examination was carried out on simplicia in
the form of powder with chloral hydrate media to
see parenchymal fragments containing oil cells,
starch grains, tracheal fragments, and other marker
characteristics possessed by simplicia.[15]
Phytochemical Screening
Tests were carried out on ganda leaf extract which
includes: Alkaloid, flavonoid, saponin, triterpenoid,
steroid, quinone, and monoterpenoids/Sesquiterpenes.
The test was based on the Farnsworth method.[16]
Check for Non-specific Parameters
Here extracts of specific gravity and drying losses
were carried out.
Determination of Moisture Content
Determination of moisture content is carried out by
distillation using toluene.[10]
Determination of Ash Total Contents
Approximately 2–3  g which had been carefully
weighed, was inserted into the silicate crucible which
had been spawned and ground, evenly distributed.
Then, it was spawned slowly until the charcoal was
used up, cooled, then weighed. Ash content was
calculated against the weight of the initial extract.[10]
Determination of Non-soluble Acid Levels
The ash obtained from the above ash content was
boiled with 25 mL dilute hydrochloric acid for 5 min,
then the insoluble part of the acid, filtered through
crucible glass or ash-free filter paper, washed with hot
water, spilled until the weight remained, then weighed.
The levels of insoluble ash in acids were calculated
against the weight of the initial extract.[10]
Specific Parameter Checking
Organoleptic extract: The organoleptic examination of
double leaf thick extract uses the senses to describe
shape, color, smell, and taste.[10]
Determination of Water Soluble Compounds
A total of 5  g of extract was macerated for 24  h
with 100  mL of chloroform LP water using clogged
flask while repeatedly shaking it for the first 6 h and
then left for 18  h. After filtering, 20  mL of filtrate
is then evaporated to dryness in a shallow flat base
cup that has been anchored; the residue is heated at
a temperature of 105ºC to a fixed weight. The level
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 Yoppi Iskandar and Resmi Mustarichie

of the water-soluble compound was calculated against Microscopic Examination Results

the initial extract.[10]
Determination of Soluble Ethanol Compounds
A total of 5 g of extract was macerated for 24 h with
100  mL ethanol (95%) using a clogged flask while
shaking repeatedly for the first 6 h and then left for 18 h,
then filtered quickly by avoiding ethanol evaporation,
then 20  mL of filtrate evaporated to dryness in a
shallow evaporator dish with a flat base, the residue
was heated at a temperature of 105ºC to a fixed weight,
then weighed. The level of the compound dissolved in
ethanol was calculated against the initial extract.
Chemical Content of Extracts
This test included the determination of essential oil
levels. The number of extracts was estimated to produce
1 mL–3 mL of essential oil add a number of extracts that
had been carefully weighed into the pumpkin connect
with the scale and cooling section. Boil the contents
of the pumpkin with the appropriate heating until the
essential oil was completely distilled and not added
again in the scale container. If the volume of essential oil
was accommodated in a sealed container, the recording
could be done with readings up to 0.1  mL, and the
volume of essential oils for every 100 g extract could
be calculated from the weight of the extract weighed.[10]
Specific Parameters
This test was carried out with TLC and GC-MS.
The results of the microscopic examination of ganda
leaf powder showed that the herb contained cover
hair, stomata, oxalate crystals, parenchyma, oil cells,
epidermis, and vessels.
Phytochemical Screening Results
Phytochemical screening was an initial description of
the compound content in one simplicia. The results of
phytochemical screening are shown in Table 1.
Huzaifa et al.[18] reported that the phytochemical
tests of Allium sativum (garlic) in its aqueous of
bulb showed the presence of flavonoids, alkaloids,
saponins, tannins, and cardiac glycosides. Abdul Kadir
et al.[19] mentioned in the phytochemical analysis of
Allium cepa L. Ethanolic extract revealed the presence
of saponins, tannins, flavonoids, and alkaloids while
steroids were not detected. Tiwari et al.,[20] showed
that phytochemical analysis of A. sativum n-butanol
fraction showed the presence of steroids, triterpenoid
saponins, and carbohydrates. Usharani et al.[21] showed
the presence of alkaloids, tannins, saponins, phenols,
flavonoids, and volatile oil that were common to all
the A. odorum ethanol extract.
Extraction
After maceration with ethanol, evaporating with rotary
evaporator followed with drying at water bath, it was
found that the viscous ethanol extract yield of ganda
leaves was 19.10 %. w/w.

TLC Specific Weight Determination

Ganda leaf extract was applied to the TLC silica gel GF The results of the determination of the specific density
254 plate using a microcapillary right at the 1 cm line of extract at the three determinations were 1.98% w/v.
from the bottom of the plate. The TLC plate was then put
into a chromatographic vessel which had been saturated Table 1: Phytochemical screening of ganda leaf extract
first with the developer of chloroform-methanol (9:1), No Chemical compounds Odorum leaves
then developed to a certain extent. After that, the ethanol extract
deadline of the developer was marked, then the spots that 1 Alkaloids +
occur were observed under 254 nm and 366 nm of UV 2 Flavonoid +
light and sprayed with the appearance of 10% sulfuric 3 Polyphenols +
4 Tannin −
acid spots in methanol, then heated at 110ºC until color
5 Quinone +
appears, then calculated the value of Rf.[17] 6 Saponins −
7 Steroids +
GC-MS 8 Triterpenoids −
This test the content of ganda leaf ethanol extracts using 9 Monoterpenoid/sesquiterpenoid +
GC-GC (Shimadzu GC-17 A; GC/MS. QP5050A). (+): Detected, (−): Not detected

Table 2: Extract parameter of ganda leaves extract

RESULTS
No Determination Results (% w/w)
Organoleptic Examination Results
1 Drying shrinkage 7.44
Ganda leaves had a width of 5–10  mm, were dense 1 Water content 12.90
and slightly sprouted, leaf length reached 20 cm, dark 2 Ash total content 5.20
3 Insoluble acid ash 0.95
green, distinctive aroma, the height of flower stalk 4 Water‑soluble compounds 10.97
reached 40 cm, white, grew with erect flower stems, 5 Soluble ethanol compounds 13.70
6 Essential oil content 0.60
solid, and square.

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 Yoppi Iskandar and Resmi Mustarichie

The results of determining the extract parameters are
shown in Table 2. All data at this table were an average
value after three determinations.
TLC Results
The developer of TLC was chloroform-methanol (9:1),
the appearance of 254 nm UV light spots, 366 nm, and
10% sulfuric acid reagent in methanol as shown in
Table 3.
GC-MS Results: The solvent was ethanol, injected
in micro size at an initial temperature of 60°C for
2–10 min until the final temperature reached 320°C for
7 min. Figure 1 and Table 4 showed GC-MS results.
DISCUSSION
In this study, the simplicia used was ganda leaves
(A. odorum L). The selection and washing of ganda
leaves were intended to remove impurities that cause
contamination and interfere with the process of
determining extract parameters.

Making extracts were done by macerating 3 × 24  h

by replacing solvents every day and occasionally
stirring so that the solvent interacts with simplicia.
The solvent replacement was intended to attract the
compound content in simplicia as much as possible.
The thick extract produced was blackish brown with
a distinctive odor. In making the extract, it was used
95% ethanol solvent, because 95% ethanol had a small
water content, thus minimizing the possibility of an
enzymatic reaction in simplicia.

The extract quality requirements consist of various

general (nonspecific) parameters, specific parameters,
and test for the chemical content of ganda leaf extract.
The non-specific parameters determined were specific
gravity, drying losses, water content, total ash content,
and acid insoluble ash content. Specific parameters
determined were the levels of water-soluble
compounds and levels of soluble ethanol compounds.
The content test carried out was the determination of
Figure 1: Gas chromatography-mass spectrometry
essential oil content, TLC, and GC-MS.
chromatogram of ganda leaves extract
The results of phytochemical screening showed
ganda leaf extract containing alkaloid compounds,
Table 3: TLC results of ganda leaves extract
flavonoids, steroids, polyphenols, quinones, and
Spots Rf UV UV 10% H2SO4 in monoterpenes/sesquiterpenes.
254 nm 366 nm MeOH
1 0.04 ‑ Blue Yellowish green From the results of non-specific parameters, it was
2 0.15 Purple ‑ ‑ known that ganda leaf extract had a specific gravity
3 0.25 Purple ‑ ‑ value of 1.84, drying losses of 7.44%, water content of
4 0.33 Purple ‑ ‑
5 0.51 ‑ Red ‑ 12.9%, total ash content of 5.2%, and acidic insoluble
6 0.68 Purple Red Light green ash content of 0.95%, whereas the results of specific
7 0.73 ‑ Red Light green parameters known as leaf extract double have a water-
8 0.81 ‑ Red Light green
9 0.91 ‑ Purple ‑ soluble compound content of 10.97% and a level of
10 0.95 ‑ Red Dark green 13.7% soluble ethanol compound. From the test of
TLC: Thin‑layer chromatography the chemical content of the extract, it could be seen

Table 4: GC‑MS of ganda leaves extract

No Rf Compounds Peak area (%)
1 25.430 2 diisooctyl esters, phthalic acid 3.3
2‑ (hexadecyloxy) ‑ 3 ‑ (octadesiloxy)
2 28.095 propyl ester, lauric acid 3.48
2‑ (1‑oxododesil) oxy ‑1,3 – propanediol ester, hexadecanoic acid
3 29.038 Phosphonic acid 43.31
4 29.583 4.31
GC‑MS: Gas chromatography‑mass spectrometry

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 Yoppi Iskandar and Resmi Mustarichie
that ganda leaf extract had an essential oil content of
0.06%, in TLC patches were produced with different
Rf values on UV 254  nm, UV 366  nm, and 10%
sulfuric acid. From the results of GC-MS, it was known
that ganda leaf extract contains 2 diisooctyl esters,
phthalic acid; 2- (hexadecyloxy) - 3 - (octadesiloxy)
propyl ester, lauric acid; 2-  (1-oxododesil) oxy-1,3-
propanedyl ester, hexadecanoic acid; phosphonic acid.
CONCLUSION
The results of this determination could be used as a
preliminary description of the quality of ganda leaf
extract quality, but the results of this determination
cannot yet be used as an A. odorum leaf extract quality
standard, because the determination of parameters
only came from one region. Further investigation of a
different area of ganda leaves is suggested in order to
obtain general parameters of ganda leaves.
ACKNOWLEDGMENT
The authors wish to thank Vivi Salfika for technical
support.
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Source of support: Nil; Conflict of interest: None Declared
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