Pharmacogn J.

2017; 9(5):683-688
A Multifaceted Journal in the field of Natural Products and Pharmacognosy Original Article
www.phcogj.com | www.journalonweb.com/pj | www.phcog.net

Indonesian Mirabilis jalapa Linn. : A Pharmacognostical and

Preliminary Phytochemical Investigations

Endang Hanani*, Rini Prastiwi, Lina Karlina

ABSTRACT
Introduction: Mirabilis jalapa Linn. is an important medicinal plant and used extensively by the people from different
countries for the treatment of several disorders. The plant was the raw material for the herb-drug product, so some
parameters identified were needed to ensure the safety, quality and efficacy of the product. Objective: The aim of this
study was to undertake pharmacognostical studies to fulfill the work required for the identification the M. jalapa plant,
which is collected from the Bogor area, Indonesia. Methods: Macroscopic and microscopic evaluation, fluorescence
standards, phytochemical screening and physicochemical parameters were carried out on the above plant. Results: The
parameters values of total ash, water soluble and acid insoluble ash were obtained 11.81, 5.06 and 0.41%, respectively.
Moisture content, alcohol, water and ether soluble extractive were found to be 12.41, 11.02, 18.63 and 7.17% respective-
ly. The results of preliminary phytochemical analysis of aqueous ethanolic extract of this drug were positive for alkaloids,
tannins, flavonoids, steroid, triterpenoids, saponin, phenols, glycosides and carbohydrate. Thin layer chromatography
(TLC) of alcoholic, chloroform and aqueous extracts showed 9, 7 and 4 spots respectively. Conclusion: The present
study on botanical pharmacognosy and TLC profile of this plant above thus provides useful information for correct iden-
tification and quality control parameters for the crude drugs, and also will be useful in making monograph of the plant.
Key words: Chromatography profile, Flourescence character, Microscopic, Nyctaginaceae, Physicochemical.

INTRODUCTION
Mirabilis jalapa Linn.(Nyctaginaceae) is a perennial some indication regarding the nature of chemical
herb and is has been called by various vernacular names compounds present in it. The pharmaceutical use
Endang Hanani*, Rini of traditionally used medicinal plants is hampered
around the world, like Gulambasa’ in Ayurveda,
Prastiwi, Lina Karlina due to the lack of standards of quality, safety and
‘Gul-abbas’ in Hindi, ‘Four o’ clock’ in English,1 and
Faculty of Pharmacy and Sciences, “pukul 4” in Indonesia.2 ‘Four o’ clock’ received the efficacy.12 The pharmacognostical and preliminary
University of Muhammadiyah name because of habit of opening in the late afternoon. phytochemistry description of a medicinal plant is
Prof. Dr. HAMKA Jl. Delima II/IV Klender, This plant is a widely used traditional medicine in many the first step towards establishing the identity and
Jakarta 13460, INDONESIA.
parts of the world for the treatment of various diseases the degree of purity of such materials and should be
Correspondence
viz. virus inhibitory activity,3 diarrhea,4 inflammation carried out before any tests are undertaken.13 This
Endang Hanani, Faculty of Pharmacy and treatment,5,6 anti-bacterial7,8 and anti-spasmodic activity.9 study is comprised the Indonesian M. jalapa Linn.
Sciences, University of Muhammadiyah
Prof. Dr. HAMKA Jl. Delima II/IV Klender,
The root is believed to be an aphrodisiac as well as
Jakarta 13460, INDONESIA. diuretic and purgative, and antioxidant activity.1,10 MATERIALS AND METHODS
Phone numbers: +62 21- 8617321/ Mirabilis jalapa has been extensively used in almost all Collection and Authentication
+62 81-188-1719; folklore remedies around the world included Indonesia The whole fresh plants material of Mirabilis jalapa
E-mail: [email protected] for treating a variety of conditions. The Indonesia Linn. were collected in the month of August –
History Traditional medicinal (“Jamu”) this plant is widely for September 2016 (from Bogor area, Indonesia), and
•  Submission Date: 29-05-2017; the treatment of diarrhea, dysentery, muscularpain,
•  Review completed: 01-06-2017; authenticated by the Research center for Biology,
•  Accepted Date: 03-06-2017
and abdominal colics, inflammation, antibacterial, Indonesian Institute of Sciences, Cibinong, Bogor,
antiviral, and antifungal functions.2 To ensure repro- Indonesia. A voucher specimen has been preserved
DOI : 10.5530/pj.2017.5.108
ducible quality of herbal products, authentication of in the Pharmacognosy laboratory, Faculty of Phar-
Article Available online
http://www.phcogj.com/v9/i5 the starting material is essential.11 This present study macy and Sciences, University of Muhammadiyah
Copyright
is concerned to characterization of different pharma- Prof. Dr. HAMKA, for further references.
© 2017 Phcog.Net. This is an open- cognostical parameters, has included here, botanical
access article distributed under the terms identification, microscopic study, powder character- Preparation of plant material
of the Creative Commons Attribution 4.0 The collected M. jalapa whole plant was washed
istics, analytical standardization, florescence study
International license.
and preliminary phytochemical screening. This pre- with tap water. The plant was cut in to small pieces
liminary study helped for standardization of the crude and air-dried thoroughly under shade (at room
drug as well as further processing of the sample with temperature) for 1 week to avoid direct loss

Cite this article : Hanani E, Prastiwi R, Karlina L. Indonesian Mirabilis jalapa Linn. : A Pharmacognostical And Preliminary
Phytochemical Investigations. Pharmacog J. 2017;9(5):683-8.

Pharmacognosy Journal, Vol 9, Issue 5, Sep-Oct, 2017 683

 Hanani et al: Pharmacognostical investigations of Mirabilis jalapa

of phyto­ constituents from sunlight. The shade dried materials Determination of acid-insoluble ashvalue
were powdered using the pulverizer and sieved up to 60 meshes, and Twenty-five (25) ml of 2N hydrochloric acid was added to the crucible
stored in a well closed glass bottle. containing the total ash, covered with a watch-glass and boiled gently
Pharmacognostical Evaluation for 5 minutes. The watch-glass was rinsed with 5ml of hot water and this
Macroscopic characteristics liquid added to the crucible. The insoluble matter was collected on an ash
less filter-paper and washed with hot water until the filtrate was neutral.
Macroscopical examination was carried out to the freshly plants material,
The filter-paper containing the insoluble matter was transferred to the
and to the powder. Morphological studies of plants material such as color,
original crucible, ignited by gradually increasing the heat to 450°C until
size, odor, taste, surface characteristic and fracture were observed based
the constant weight was obtained. The percentage of acid-insoluble ash
on the description given in Evans WC14 and Indonesian Herb Pharmaco-
was calculated with reference to the air dried drugs.
poeia.15 Organoleptic characters were observed, noted and photographs
were taken in the original environment. Water extractive value
Microscopic characteristics Separately place about 10.0 g (accurately weigh) of whole plant powder
of the M. jalapa, in an accurately weighed glass stopper conical flask.
Mature, fresh and healthy the different plant parts as leaf, stem, root,
Then 100 ml of distilled water was added to the flask and weighed to
etc. and the powder were used for microscopic evaluation.The trans-
obtain the total weight including the flask. The contents was macerated,
verse sections of the different plant parts were prepared by free hand,
shaken well during the first 6 hrs and allowed to stand for 18 hours.The
it was put between the pith and fine sections were cut with the help of a
solution filtered rapidly through a dry filter.Theflask was readjusted to
sharp razor. The sections so obtained were cleared using chloral hydrate
the original total weight with distilled water. Twenty-five ml of the
solution.Preparing for powder microscopy, small amount of whole plant
filtrate was transferred to an accurately weighed, tarred flat-bottomed
material powder was placed on a glass slide containing one to two drops
dish and evaporated to dryness on a water-bath. Finally, it was dried
of chloral hydrate solution. After placing the cover slip, it was warmed
at 105°C until the constant weight was obtained. The percentage of the
gently on spirit lamp to remove air bubbles and then observed under the
value of water extractive was calculated with reference to the air-dried
microscope. Different tissues were observed under the microscope and
drug.
were photographed.16,17
Ethanol extractive value
Physicochemical Evaluation
Same procedure as water extractive valuewas followed using ethanolin-
Physicochemical characteristics such as moisture content,total ash, water
stead of distilled water to determine extractable matter in ethanol. The
soluble and acid insoluble ash tests were performed according to the
ethanol extractive value was calculated with reference to the air-dried
official methods and the WHO guidelines on the quality control methods
drug.
for medicinal plant materials.13,1 The determination of the extractive
values (alcohol, water and ether soluble)of the powdered drug was carried Ether extractive value
out according to the procedure described in Indonesian Herb Pharmaco- Same procedure as water extractive valuewas followed using ether to
poeia, 2008.15 Fluorescence characters of powdered and extracts material determine extractable matter in ether. The ether extractive value was
with different chemical reagents were determined under ordinary and calculated with reference to the air-dried drug.
ultraviolet light.
Fluorescence Characters
Moisture content Fluorescence characters of powdered drug material were sieved through
About 2 g of the air dried powdered drug was weighed in a watch glass, 60 mesh and observations with different chemical reagents.These were
kept in hot air oven at 1050C and dried for a period (about 30 minutes) observed under visible light, short and long ultraviolet (254 and 366 nm).
until constant weight was obtained. Weight loss on drying was noted and The reagents were methanol, 2N hydrochloric acid, 50% sulphuric acid,
difference in weight gives the moisture content of powdered drug. Total 50% nitric acid and 5% potassium and sodium hydroxide.18,19 The intensity
moisture content of the drug was noted. of the coloration determines the abundance of the compound present.
Determination of total ash value Phytochemical Screening
Two grams of the powdered material was placed in a silica crucible which Weighed 25 g of plant material powdered was extracted in a reflux
previously was ignited and tarred crucible accurately weighed.The apparatus with 250 ml methanol for 30 minutes. The extracts were filtered
powdered material was spread as a fine even layer at the bottom of the and evaporated to dryness under reduced pressure and controlled temper-
crucible. The crucible was incinerated until the temperature about 450oC, ature (40-50ºC). The extracts were subjected to preliminary phytochemical
and a white material was obtained, that is indicating the absence of investigation for the detection of following compounds; carbohydrates,
carbon. The crucible was cooled and weighed. The procedure was protein, steroids and terpenoids,glycosides, flavonoids, alkaloids, tannins,
repeated until the constant weights. The percentage of the total ash was phenolic compounds anthraquinones and saponins, as per the stan-
calculated with reference to the air dried powdered sample. dard procedures described by Indonesian Herb Pharmacopoeia and
Harborne.15,20 The presence of phyto-constituents from the above plants
Determination of water soluble ash value
was presented in the Table 3.
The total ash obtained was boiled with 25 ml of water for five min. The
insoluble matter was collected on a ash less filter paper & and washed Chromatographic Profile
with hot water. The insoluble ash was transferred into pre-weighed silica The powder material (10 g) was subjected it to successive Soxhlet extraction
crucible, ignited for 15 min at a temperature not exceeding 450oC, cooled with 100 ml solvents: hexane, dichloromethane (DCM) and methanol
and weighed.The procedure was repeated to get the constant weight. The respectively. The extraction was continued until the solvent became
weight of the insoluble matter was subtracted from the weight of total colorless. The 3 extracts were concentrated using a rotary evaporator
ash. The percentage of water soluble ash was calculated with reference to then analyzed by TLC using silica gel 60 F254 TLC plates for the chro-
the air-dried sample drug. matographic profile.Each extract was faintly dissolved in methanol and

684 Pharmacognosy Journal, Vol 9, Issue 5, Sep-Oct, 2017

 Hanani et al: Pharmacognostical investigations of Mirabilis jalapa

capillary tubes were used to uniformly apply the dissolved samples on 2.5-3.5 cm length. Leaves are green, slightly bitter and having character-
the plates and allowed to dry. The mobile phases for the plates developed istic odor. These are ovate shape, pinnatified, acuminate apex, crenate,
were hexane –DCM (6:4), chloroform- methanol (9:1), ethyl acetate – cordate base and 5–11cm in length, 4–7 cm in width. The plants have
methanol (7:3). The plates were dried and observed under visible light black carrot shaped tubers that can be a foot or more length, and wrinkled
and ultraviolet light 366nm, and by spraying with 10% sulfuric acid black obovoid fruits (Figure 1).
followed by heating at 105oC for 5-10minutes in an oven.21The retention
Microscopic Studies
factor (Rf) value was calculated by using this formula:
Transverse section of M. jalapa Linn leaf shows presence of multicellular
Rf = Distance moved by the solute (compound) / Distanced moved by trichomes on both surfaces. The upper and lower epidermis consists of
the solvent front. oval shaped parenchyma cells in single layer and also absence of stomata.
The vascular bundles are composed of both xylem and phloem in
collateral open arrangements. M. jalapa Linn. leaf also show the presence
RESULTS
of cubuoidal and raphides calcium oxalate crystal (Figure 2A, 3A).
Macroscopic Studies Transverse section of M. jalapa steam show much trichomes, single cell
Mirabilis jalapa Linn. (Nyctaginaceae) is a perennial herb that reaches a of epidermis, collenchyma and some layer of parenchyma cells (Figure 2B).
height of 50-100 cm, and a popular ornamental plant grown worldwide Transverse section of the root show the cubuoidal calcium oxalate crystal
for the beauty of its flowers. They have numerous branches and opposite, and raphides calcium oxalate crystal (Figure 2C), and Figure 3A showed
and the flowers are borne singly or in clusters, and can be red, magenta, also the cubuoidaland raphides calcium oxalate crystal. Fragment of
pink or yellowish, have light fragrance characteristic taste. Individual stigma, anther and vascular bundles were showed in Figure 3B, 3C and
flowers are trumpet shaped, about an inch across at the end and 3D respectively.

Figure 1: A. Tuber, B. Branch, C. Seeds, D. Fresh leaf, and E. Flower of M. jalapa.

Figure 2: A. Transverse section of leaf: a. trichome; b. raphides calcium oxalate crystal, c. sclereid cell, d. cubuoidal calcium oxalate
crystal. B. Transverse section of steam: a. epidermis hair, b. collenchyma, c. pith, d. endodermis, e. phloem, f. xylem, C. Transverse section
of root: a. endodermis, b. cubuoidal calcium oxalate crystal, c. cambium, d. raphides calcium oxalate crystal.

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 Hanani et al: Pharmacognostical investigations of Mirabilis jalapa

Figure 3: A. Raphides and cubuoidal calcium oxalate crystal, B. Portion of stigma; C. Fragment of anthers with oil gland, D. Vascular
bundles

Physicochemical characteristics Phytochemical Screening

The physico-chemical characteristics such loss on drying, ash values, PreliminaryPhytochemical screening determination of ethanolic extract
water, alcohol and ether soluble extractive, were given in the Table 1. by some chemical test was carried out. The results were positive for alka-
loids, tannins, flavonoids, glycosides, phenols, steroid, triterpenoids and
Fluorescence Characters
saponin, carbohydrate (Table 3).
Fluorescence characters of powdered material were analyzed under
visible and ultraviolet light (254 and 366 nm), which signifies their char- Chromatographic Profile
acteristics (Table 2). Thin layer chromatographic profile evaluated of M.jalapa leaves extracts
(hexane, DCM, ethanol) constituted different colored phytochemical
Table 1: Physicochemical Parameter of Powder of M. jalapa compounds with different Rf value. The Rf values were calculated for
Sl.No Parameters Average (%w/w) the optimum solvent system revealed the presence of promising spots as
shown in Table 4.
1 Moisture content
Loss on drying 12.41% ± 0.005 DISCUSSION
2 Ash Value
Macroscopical and microscopical characters will help in the identifica-
a. Total ash 11.81% ± 0.001
tion of right variety and search for adulterants. The morphological char-
b. Water soluble ash 5.06% ± 0.001 acters of different parts of the plant can serve as diagnostic parameters.
c. Acid insoluble ash 0.41% ± 0.001 Microscopic evaluation is one of the simplest and cheapest methods for
3 Extractive values establishing the correct identification of the source of the materials. In
a. Alcohol soluble 11.02% ± 0.007 this microscopical character of M. jalapa leaves show the presence of
b. Water soluble 18.63% ± 0.007
c. Ether soluble 7.17% ± 0.007 Table 3: Phytochemical Screening of Plant Extracts of M. jalapa
Phytoconstituens Test Performed /reagents Results
Table 2: Fluorescence Analysis of Powder and Ethanolic Extract Carbohydrates Fehling +
of M.jalapa
Molisch +
Treatment Normal light UV (254 nm) UV (366 nm) Proteins Biuret -
Powder Ninhydrin -
+Water Green Light yellow Light yellow Alkaloids Dragendorff +
+ KOH Brown Dark brown Brown Mayer +
Bouchardat +
+NaOH Greenish brown Yellowish brown Brown
Anthraquinones Borntrager -
+H2SO4 Light brown Bright yellow Light brown
Flavonoids Shinoda +
+HCl Light brown Bright yellow Light yellow Ammonia +
+HNO3 Yellowish brown Greenish brown Greenish brown Glycosides Molisch +
Alcohol Extract 70% Fehling +
+Water Brown Yellow Yellow Phenols Ferric chloride +
Saponins Foam +
+ KOH Yellowish brown Brown Light Brown
Steroids - terpenoids Liebermann +
+NaOH Greenish brown Yellowish brown Greenish brown BurchardSalkowski +
+H2SO4 Light brown Bright yellow Yellow
Tannins Gelatin +
+HCl Brown Bright yellow Yellow Lead acetate +
+HNO3 Light brown Brown Light brown
Description: (+) = present; (-) =Absent

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 Hanani et al: Pharmacognostical investigations of Mirabilis jalapa

Table 4: Chromatographic Profile of Different Extracts of M.jalapa Leaves

Extract Solvent System Total Spraying reagent Rf Value
Spots (10% H2SO4)
Hexane Hexane - DCM 6:4. 9 Light brown, light grey, yellow, light grey, light yellow, light grey, light yellow, 0.08, 0.17, 0.28, 0.35, 0.50, 0.60,
light yellow, yellow 0.75, 0.86, 0.92
DCM Choroform – 7 Light brown, light grey, grey, light yellow, yellow, light blue, light yellow. 0.05, 0.15, 0.46, 0.60, 0.68, 0.75,
methanol 9:1. 0.90
Ethanol Ethyl acetate – 4 Light brown, light yellow, yellow, light yellow 0.05, 0.31, 0.41,
ethanol 7:3 0.90.

cubuoidal22,23 and raphides needle in shape of calcium oxalate crystal in CONFLICT OF INTEREST
Indonesian drug. Drying plays a very important role in the quality as
well as purity of the material. Moisture was not so high as to facilitate Authors declare no conflict of interest.
activation of enzymes and gives suitable condition, to the proliferation
of microorganisms, and is an inevitable component of crude drugs,
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 Hanani et al: Pharmacognostical investigations of Mirabilis jalapa

GRAPHICAL ABSTRACT HIGHLIGHTS OF PAPER

•  Mirabilis jalapa Linn. has been called by various vernacular names around the
world, like Gulambasa’ in Ayurveda, ‘Gul-abbas’ in Hindi, ‘Four o’ clock’ in
English,1 and “pukul 4” in Indonesia.
•  This plant is used for treatment of various diseases viz. virus inhibitory activ-
ity, diarrhea, inflammation treatment, anti-bacterial and anti-spasmodic activity.
•  M.jalapa origin from Indonesia, India and South India showed the same phyt-
ocons-tituens compounds.
•  This is the first report of Indonesian M. jalapa Linn. on the pharmacognostical
studies, and provides useful information for quality control parameters for the
crude drugs.

AUTHOR PROFILE
Endang Hanani: Lecturer and researcher at Faculty of Pharmacy and Sciences, University of Muhammadiyah Prof. Dr. HAMKA, Klender, Ja-
karta 13460, Indonesia. Also as Professor Pharmacognosy and Phytochemistry.

Rini Prastiwi: Lecturer and researcher at Faculty of Pharmacy and Sciences, University of Muhammadiyah Prof. Dr. HAMKA, Klender, Jakarta
13460, Indonesia.

Lina Karlina: Graduate student at Faculty of Pharmacy and Sciences, University of Muhammadiyah Prof. Dr. HAMKA, Klender, Jakarta 13460,
Indonesia.

Cite this article : Hanani E, Prastiwi R, Karlina L. Indonesian Mirabilis jalapa Linn. A Pharmacognostical And Preliminary Phytochemical Investigations. Pharmacog J.
2017;9(5):683-8.

688 Pharmacognosy Journal, Vol 9, Issue 5, Sep-Oct, 2017