Standardization Raw Material and Ethanolic Extract of
Andrographidis Herba (Andrographis Paniculata Nees) from District
of Bogor and Tawangmangu
Sukardiman, Rakhmawati, Alvi Prabowo , Lusiana Arifianti
Department of Pharmacognosy and phytochemical, Faculty of Pharmacy, Airlangga University, Surabaya, Indonesia
Keyword: Standardization, raw material, ethanolic extract, Andrographis paniculata Nees, Andrographis herbs
Abstract: Sambiloto (Andrographis paniculata Nees) has a potency to be developed in phyto-pharmaceutical product,
especially in this study focused on supporting anti-hyperglycemic research. This requires standardization
can affect both the raw material or extract qualities that have been obtained based on the Indonesian Herbal
Pharmacopeia method. Raw material for this study has been obtained from two traditional drug
development institutions, there are BALITRO from Bogor and BPTO from Tawangmangu
1 INTRODUCTION
Sambiloto (Andrographis paniculata Nees) is one of
medicinal plant that it has long existed and used as
ingredient of traditional medicine. Sambiloto has
bitter taste, contains flavonoids, tannins, and
saponins [1].
Ethanolic extract of sambiloto can restore blood
glucose level into normal in induced-rats, it
indicated as anti-hyperglicemic activity [2]. Another
study showed that combination curcumin and
andrographolide (3:1) have role in reducing blood
glucose level in rats by 47.945% was preprandial
and 74.159% was postprandial [3]. The study of
sambiloto as anti-hyperglicemic also showed in the
mixture formula such as sambiloto and mahoni
reduced blood glucose level in mice by 250.80 mg/dl
[4].
Many studies have proven the efficacy of
sambiloto as anti-diabetic agent and raises potential
to be developed into phyto-pharmaceutical product
as anti-diabetic. Therefore, it is necessary to do
standardization. It can affect quality of both extract
and raw material by methods that have been
established by Ministry of Health through Indonesia
Herbal Pharmacopeia. It is a guarantee for safety,
effectiveness, and especially quality of plant
material, raw material and ethanolic extract that
developed into phyto-pharmaceutical preparation.
2 MATERIAL AND METHODS
2.1. Plant Material
In this research, the raw material was obtained from
two different districts, there are Bogor (BALITRO,
Balai Penelitian Tanaman Obat dan Aromatik) and
Tawangmangu (BPTO, Balai Besar Penelitian dan
Pengembangan Tanaman Obat dan Obat
Tradisional). They prepared in dried raw material.
The ethanolic extract was extracted by maceration
method with 96% ethanol for 24 hours and
performed up to three times [5]. The mixture extract
was concentrate using rotary evaporator.
2.2. Determination of Specific and Non-
specific Parameters of Dried Raw
Material and Ethanolic Extract of
Sambiloto
Determination of standard parameter was done using
Ministry of Health Guidelines through Indonesian
Herbal Pharmacopeia [6]. Specific parameters of
raw material and extract were macroscopic,
macroscopic, organoleptic, water-soluble extractive,
ethanol soluble extractive, and andrographolide
assay in raw material as well as ethanolic extract.
While non-specific parameters encompassed loss on
Sukardiman, ., Rakhmawati, ., Prabowo, A. and Arifianti, L.
Standardization Raw Material and Ethanolic Extract of Andrographidis Herba (Andrographis paniculata Nees) from District of Bogor and Tawangmangu.
DOI: 10.5220/0009846000002406
In Proceedings of BROMO Conference (BROMO 2018) - Symposium on Natural Product and Biodiversity, page 1
ISBN: 978-989-758-347-6
Copyright
c
2022 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
1
drying, total ash content, water content, and acid-
insoluble ash content. In this case, the assay of
andrographolide was based on previous study to
prove andrographolide was major compound that
had anti-hyperglicemic activity.
2.3. Determination of Andrographolide
Content in Raw Material and
Ethanolic Extract
Andrographolide content in raw material and extract
determined based on the sambiloto monograph in the
Indonesian Herbal Pharmacopeia. Andrographolide
used to make calibration curve. Concentration of the
andrographolide solution was 0.8 and 0.125 mg/ml.
Andrographolide solution at 0.8 mg/ml was dropped
with 1.2 and 3 µl, whereas the solution at 0.125
mg/ml was dropped with 2.4 and 6 µl of volume.
The extract solution and ethanol soluble extractive
of raw material were made at 1 mg/ml.
The thin layer chromatography system was
silica-gel as stationary phase and the mobile phase
system of chloroform:methanol (9:1) [7]. Then,
sample eluted with mobile phase until the spot
moved up to 8 cm. The spot was measured with TLC
densitometry with lambda 310 nm.
3 RESULT AND DISCUSSION
Bogor district is representative city with high rainfall
(4.086 mm/year). It has altitude at 190 330 m
above of the sea level. The soil characteristic in the
Bogor is yellowish-red latosol. Whereas
Tawangmangu locations has altitude at 1236 m
above of the sea level and the soil condition is
dominated by brown litosol. Tawangmangu district
was represented with high rainfall (3326 mm/year).
The difference environment may influence
component in the andrographis herbs.
3.1. Macroscopic Study
Figure 1: Dry herbs of sambiloto (A) from Bogor and (B)
from Tawangmangu
3.2. Microscopic Study
There was eight identifier fragments in each
sambiloto from two different places. Mesophyll
tissues with leaves bone were identified (fig. 2A).
Another fragment was discovered as stomata with
diacytic type (fig. 2B) and xylem with dot type (fig.
2C) and ladder type (fig. 2D). Cystolith was
discovered in the epidermal wall (fig. 2E). The
trichoma was found in the leaves (fig. 2F) and
chollenchyma was often found in the stems (fig.
2G).
Table 1: Result of phytochemical identification of raw
material (dried herbs) and ethanolic extract of
Andrographis panciulata Nees
3.3. Physicochemical Identification
The results of physicochemical identification were
showed in the table 1. They showed that several
parameters such as water-soluble extractive loss on
drying, and water content of extract from Bogor was
higher than Tawangmangu, oppositely that ethanol
soluble extractive of sample and insoluble acid ash
from Tawangmangu higher than Bogor. Both ash
content from Bogor or Tawangmangu showed
higher than Indonesian Herbal Pharmacopeia
Phytochemical
parameters
Region
Sample form
Mean ± SD
(n=3)
1. Water content
(%)
Bogor
ethanol extract
12.28 ±
0.43
Tawangmangu
ethanol extract
9.97 ±
0.01
2. Total ash (%)
Bogor
ethanol extract
3.12 ±
0.34
dried herbs
11.81 ±
0.06
Tawangmangu
ethanol extract
7.45 ±
1.45
dried herbs
11.34 ±
0.52
3. Acid insoluble
ash (%)
Bogor
ethanol extract
0.03 ±
0.01
dried herbs
0.09 ±
0.08
Tawangmangu
ethanol extract
0.04 ±
0.01
dried herbs
0.53 ±
0.02
4. Loss on drying
(%)
Bogor
dried herbs
8.66 ±
0.26
Tawangmangu
dried herbs
7.91 ±
0.18
5. Ethanol soluble
extractive (%)
Bogor
dried herbs
9.98 ±
0.09
Tawangmangu
dried herbs
10.29 ±
0.33
6. Water soluble
extractive (%)
Bogor
dried herbs
17.41 ±
0.43
Tawangmangu
dried herbs
16.16 ±
0.99
7.Andrographolide
content (%)
Bogor
ethanol extract
23.76 ±
1.06
dried herbs
2.25 ±
0.02
Tawangmangu
ethanol extract
15.61 ±
0.88
dried herbs
1.95 ±
0.02
A
B
BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity
2
requirement. The water content should not more
than 10% because it may affect stability during
storage and production caused by enzymatic activity.
The total ash content was higher than Indonesian
Herbal Pharmacopeia requirement either from Bogor
or Tawangmangu. The contaminations of total ash of
the both samples were exceeded from limit
requirement of Indonesia Herbal Pharmacopeia, but
insoluble acid ash of both samples still at allowed
level.
Figure 2 : Identifier fragments for sambiloto in the left side are from Bogor and right side are from Tawangmangu
3.4. Determination of the
Andrographolide Content
Andrographolide (C
20
H
30
O
5
) is a major diterpenoid
in A. paniculata that presence about 4%, 0.8~1.2%
and 0.5~6% in dried whole plant, stem and leaf
extracts, respectively [2,3,5]. Their spot appeared
at 0.37 of Rf value. The sample from Bogor had
andrographolide higher than from Tawangmangu,
not only as raw material but also as extract.
Andrographolide content in ethanolic extract from
Bogor was 23.76 ± 1.06 and from Tawangmangu
was 15.61 ± 0.88, whereas andrographolide
content in dried herbs was 2.25 ± 0.02 from Bogor
district and 1.95 ± 0.02 from Tawangmangu
district.
In conclusion, the results of standard
parameters from Bogor and Tawangmangu
allowed at level of Indonesia Herbal
Pharmacopeia, except total ash content of ethanolic
extract and dried herbs and water content of
ethanolic extract sambiloto from Bogor was
exceeded from Indonesian Herbal Pharmacopeia
requirements.
REFERENCES
Adelyna. Penelusuran senyawa bioaktif sambiloto
(Andrographis paniculata Nees) dengan metode
Brine Shrimp Lethality Test (BSLT). Bogor: Fateta-
Bogor Agricultural University; 1999.
Yu BC, Hung CR, Chen WC, Cheng JT:
Antihyperglycemic effect of andrographolide in
streptozotocin induced diabetic rats. Planta Med.
2003, 69: 1075-1079.2
Cheung HY, Cheung CS, Kong CK: Determination of
bioactive diterpenoids from Andrographis
paniculata by micellar electrokinetic
chromatography. J Chromatogr A 2001, 930(1-
2):171-176.
Sukardiman, Riza NF, Rakhmawati, Studiawan H,
Santosa MH, Rahman A. Hypoglicemic activity of
96% Ethanolic Extract of Andrographis paniculata
Nees. and Swietenia mahagoni Jacq. Combination.
E-Journal Planta Husada 2012; 1:1-9.
Pholphana N, Rangkadilok N, Thongnest S, Ruchirawat
S, Ruchirawat M,Satayavivad J: Determination and
variation of three active diterpenoids in
Andrographis paniculata (Burm.f.) Nees. Phytochem
Anal 2004, 15(6):365-371.
Kementrian Kesehatan Republik Indonesia. Farmakope
Herbal Indonesia Edisi pertama. Jakarta: Kementrian
Kesehatan RI;2008.
Safitri. Penetapan parameter standar simplisia dan
ekstrak etanol herba sambiloto (Andrographis
2A
2A
2B
2B
2C
2C
2D
2D
2G
2G
2E
2E
2F
2F
Standardization Raw Material and Ethanolic Extract of Andrographidis Herba (Andrographis paniculata Nees) from District of Bogor and
Tawangmangu
3
paniculata Nees). Surabaya: Airlangga University;
2007.
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