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

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

BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity

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

) 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.

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BROMO 2018 - Bromo Conference, Symposium on Natural Products and Biodiversity